Feed additive compositions

ABSTRACT

The present invention provides feed compositions comprising a basal animal diet supplemented with direct fed microbial (DFM) and at least one of a formulated yeast, a  capsicum  product, and an antimicrobial clay. The feed compositions improve growth, general health, intestinal health, and reduce microbial pathogens in non-human animals fed the feed compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/587,860 filed Nov. 17, 2017, the entire disclosure ofwhich is incorporated herein by reference.

FIELD OF THE INVENTION

The present disclosure generally relates to feed additive compositions,methods for improving growth, improving health, improving intestinalhealth, and the reduction of microbial pathogens in non-human animals.

BACKGROUND OF THE INVENTION

Animal husbandry is a branch of agriculture concerned with raising,breeding, and day to day care of animals. Farmers engaged in this branchof agriculture produce meat, eggs, milk, and other products for theconsumer. Additionally, these farmers are engaged in breeding andraising a wide variety of animals.

Farmers engaged in animal husbandry face many challenges. The demand forfood and food products from animal husbandry is anticipated to increasesignificantly as the population is growing. Also, with the increase inpopulation, increased demands on land, water, and energy resources arebeing realized. Global environmental challenges, including globalclimate changes and the growing threat of disease transmission to andfrom agricultural animals, add further challenges. Therefore, farmersneed to become more efficient, produce the products at a higher rate,and raise livestock and poultry at an increased rate to meet marketchallenges. Additionally, farmers need a low cost method to producethese products since the profit margin in these areas can be quite low.

In the past, farmers used antibiotics and hormones to not only controldiseases and pathogens but also to promote growth in livestock andpoultry. Due to changes in consumer sentiment towards the use ofantibiotics, there is an increase of production that is reared withoutthe use of antibiotics.

To meet these challenges, what is needed is a feed additive which isgenerally low cost but also promotes health, growth, and reduces thelevels of pathogens in animals without having to use antibiotics andhormones.

SUMMARY OF THE INVENTION

In one aspect, the present disclosure provides a feed composition fornon-human animals. The composition comprises a basal animal dietsupplemented with a combination of active ingredients selected fromcapsicum product and a DFM; antimicrobial clay and a DFM; formulatedyeast and a DFM; capsicum product, formulated yeast, and a DFM, andcapsicum product and formulated yeast.

The DFM may be Bacillus licheniformis. When the the DFM is Bacilluslicheniformis, the Bacillus licheniformis in the feed composition may beat a concentration of about 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.

The capsicum product in the feed composition may be at a concentrationof about 0.1 lb/ton to about 0.5 lb/ton feed. The capsicum product maybe selected from the group consisting of a capsaicinoid, a vanilloid,capsicum, macerated hot peppers, ground hot peppers, hot pepper extract,capsaicin-containing plant materials, encapsulated ground peppers, acoated capsaicin product, and combinations of any thereof. In someembodiments, the capsicum product is encapsulated ground peppers. Whenthe capsicum product is encapsulated ground peppers, the capsicumproduct comprises about 45% to about 55% ground peppers and/or comprisesabout 0.4% to about 0.6% capsaicinoids.

The formulated yeast may comprise about 80-100% yeast extract. When theformulated yeast comprises yeast extract, the formulated yeast in thefeed composition may be at a concentration of about 0.3 lb/ton of feedto about 0.7 lb/ton of feed. The antimicrobial clay may be mined clay.When the antimicrobial clay is mined clay, the clay in the feedcomposition is at a concentration of about 2 to about 6 lb/ton feed.

In some embodiments, the composition comprises formulated yeast,capsicum product, and DFM. In an alternative of the embodiments, the DFMis Bacillus licheniformis. In an alternative of the embodiments whereinthe DFM is Bacillus licheniformis, the formulated yeast in the feedcomposition is at a concentration of about 0.3 lb/ton to about 0.7lb/ton feed, the capsicum product in the feed composition is at aconcentration of about 0.1 lb/ton to about 0.5 lb/ton feed, and theBacillus licheniformis in the feed composition is at a concentration ofabout 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.

In other embodiments, the composition comprises capsicum product andDFM. In an alternative of the embodiments, the DFM is Bacilluslicheniformis. In an alternative of the embodiments wherein the DFM isBacillus licheniformis, the capsicum product in the feed composition isat a concentration of about 0.1 lb/ton to about 0.5 lb/ton feed and theBacillus licheniformis in the feed composition is at a concentration ofabout 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.

In yet other embodiments, the composition comprises antimicrobial clayand DFM. In an alternative of the embodiments, the DFM is Bacilluslicheniformis. In an alternative of the embodiments wherein the DFM isBacillus licheniformis, the antimicrobial clay in the feed compositionis at a concentration of about 2 lb/ton to about 6 lb/ton feed and theBacillus licheniformis in the feed composition is at a concentration ofabout 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.

In other embodiments, the composition comprises formulated yeast, andDFM. In an alternative of the embodiments, the DFM is Bacilluslicheniformis. In an alternative of the embodiments wherein the DFM isBacillus licheniformis, the formulated yeast in the feed composition isat a concentration of about 0.3 lb/ton to about 0.7 lb/ton feed, and theBacillus licheniformis in the feed composition is at a concentration ofabout 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.

In additional embodiments, the composition comprises formulated yeast,and capsicum product. In an alternative of the embodiments, theformulated yeast in the feed composition is at a concentration of about0.3 lb/ton to about 0.7 lb/ton feed, and the capsicum product in thefeed composition is at a concentration of about 0.1 lb/ton to about 0.5lb/ton feed.

In other aspects, the present disclosure provides a feed composition fornon-human animals comprising a basal animal diet supplemented withcapsicum product and Bacillus licheniformis; a feed composition fornon-human animals comprising a basal animal diet supplemented withantimicrobial clay and Bacillus licheniformis; a feed composition fornon-human animals comprising a basal animal diet supplemented withformulated yeast and Bacillus licheniformis; a feed composition fornon-human animals comprising a basal animal diet supplemented withcapsicum product, formulated yeast, and Bacillus licheniformis; or afeed composition for non-human animals comprising a basal animal dietsupplemented with capsicum product and formulated yeast.

In another aspect, the present disclosure provides a method forimproving health in non-human animals, the method comprising feeding thenon-human animal the feed composition described above. Improving thehealth of a non-human animal may be selected from reducing incidence ofdiarrhea, reducing number of days of diarrhea, decreasing mortality,improving intestinal health, reducing microbial pathogens in theintestinal tract of the animal, decreasing cytokine panel measuringTNF-alpha, and combinations thereof. Further, the non-human animal maybe a chicken. When the non-human animal is a chicken, improving thehealth of the chicken comprises reducing the impact of C. perfringens inbroilers.

In yet another aspect, the present disclosure provides a method forimproving performance of non-human animals, the method comprisingfeeding the non-human animal the feed composition described above.Improving performance may be selected from increase in body weight gain,feed conversion rate (FCR), feed intake, average daily weight gain(ADG), an increase in average daily food intake (ADFI), an improvedoverall body weight, and ratio of F/G. The non-human animal may be achicken.

In yet another aspect, the present disclosure provides method forimproving quality of a product derived from a non-human animal, themethod comprising feeding the non-human animal the feed compositiondescribed above. Improving the quality of a product derived from anon-human animal may be selected from from improving breast meat yield,reducing product variability, reducing impact on consumer appeal due topoor color, poor texture, and drip loss in retail display, reducingorganoleptic qualities in cooked chicken, including incidence of woodybreast. The non-human animal may be a chicken.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure is based on the discovery that administering tonon-human animals a feed composition comprising at least two activeingredients selected from a direct fed microbial (DFM), a formulatedyeast, a capsicum product, and an antimicrobial clay increases animalproduction, improves animal health, and/or improves performance. Assuch, the instant invention is directed generally to basal animal dietssupplemented with at least two active ingredients selected from a directfed microbial (DFM), a formulated yeast, a capsicum product, and anantimicrobial clay, and methods of using the feed compositions. Forinstance, feed compositions may be used to improve the health of anon-human animal. Additionally, feed compositions may be administered toanimals to improve mortality, growth performance and feed conversion,and improve the quality of products from animals.

(I) Feed Compositions

One aspect of the present disclosure encompasses feed compositions fornon-human animals. The compositions comprise a basal animal dietsupplemented with at least two active ingredients selected from a directfed microbial (DFM), a formulated yeast, a capsicum product, and anantimicrobial clay. In some embodiments, feed compositions comprise abasal animal diet supplemented with capsicum product and a DFM. In otherembodiments, feed compositions comprise a basal animal diet supplementedwith antimicrobial clay and a DFM. In yet other embodiments, feedcompositions comprise a basal animal diet supplemented with formulatedyeast and a DFM. In additional embodiments, feed compositions comprise abasal animal diet supplemented with capsicum product, formulated yeast,and a DFM. In some embodiments, feed compositions comprise a basalanimal diet supplemented with capsicum product and formulated yeast.

(a) Direct Fed Microbial

As used herein, the term “direct fed microbial” (DFM) is used to referto a probiotic or prebiotic living microorganisms that provide healthbenefits when ingested by an animal. DFMs may include yeast, bacteria,and combinations thereof.

By way of non-limiting example, yeast DFMs may comprise Saccharomycesbisporus, Saccharomyces boulardii, Saccharomyces cerevisiae,Saccharomyces capsularis, Saccharomyces delbrueckii, Saccharomycesfermentati, Saccharomyces lugwigii, Saccharomyces microellipsoides,Saccharomyces pastorianus, Saccharomyces rosei, Candida albicans,Candida cloaceae, Candida tropicalis, Candida utilis, Geotrichumcandidum, Hansenula americana, Hansenula anomala, Hansenula wingei, andAspergillus oryzae.

Non-limiting examples of bacterial DFMs include Lactobacillusacidophilus, Bifedobact thermophilum, Bifedobat longhum, Streptococcusfaecium, Bacillus pumilus, Bacillus subtilis, Bacillus licheniformis,Lactobacillus acidophilus, Lactobacillus casei, Enterococcus faecium,Bifidobacterium bifidium, Propionibacterium acidipropionici,Propionibacteriium freudenreichii, and Bifidobacterium pscudolongum.

The amount of DFM in a feed composition can and will vary depending onthe DFM, the type of non-human animal that will be administered the feedcomposition, the body weight, sex, and medical condition of thenon-human animal among other variables, and can be determinedexperimentally. Preferably, a DFM suitable for feed compositions of thedisclosure is Bacillus Licheniformis. When a DFM of a feed compositionis Bacillus Licheniformis, the concentration of Bacillus Licheniformisin the feed composition may range from about 1×10¹ cfu/g of feed toabout 1×10¹⁰ cfu/g of feed, from about 1×10¹ cfu/g of feed to about1×10⁶ cfu/g of feed, from about 1×10⁵ cfu/g of feed to about 1×10¹⁰cfu/g of feed, or from about 1×10³ cfu/g of feed to about 1×10⁸ cfu/g offeed. Preferably, the concentration of Bacillus licheniformis is about1×10⁴ to about 1×10⁶ cfu/g feed.

(b) Formulated Yeast

As used herein, the formulated yeast product may comprise a combinationof Saccharomyces cerevisiae yeast extract representing approximately25-100% of the total formulated yeast product by weight, hydrolyzedyeast representing approximately 0-40% of the total formulated yeastproduct by weight, a yeast culture representing approximately 0-50% ofthe total formulated yeast product by weight. The formulated yeast mayalso comprise limestone representing approximately 0-50% of the totalformulated yeast product by weight.

The formulated yeast may be any yeast provided the yeast is generallyregarded as safe for use in food or medical applications. Non-limitingexamples of formulated yeast-derived products may include yeast cellwall derived components such as β-glucans, arabinoxylan isomaltose,agarooligosaccharides, lactosucrose, cyclodextrins, lactose,fructooligosaccharides, laminariheptaose, lactulose,β-galactooligosaccharides, mannanoligosaccharides, raffinose, stachyose,oligofructose, glucosyl sucrose, sucrose thermal oligosaccharide,isomalturose, caramel, inulin, and xylooligosaccharides. In anembodiment, the formulated yeast may be β-glucans and/ormannanoligosaccharides. Sources for yeast cell wall derived componentsinclude Saccharomyces bisporus, Saccharomyces boulardii, Saccharomycescerevisiae, Saccharomyces capsularis, Saccharomyces delbrueckii,Saccharomyces fermentati, Saccharomyces lugwigii, Saccharomycesmicroellipsoides, Saccharomyces pastorianus, Saccharomyces rosei,Candida albicans, Candida cloaceae, Candida tropicalis, Candida utilis,Geotrichum candidum, Hansenula americana, Hansenula anomala, Hansenulawingei, and Aspergillus oryzae.

The formulated yeast may also include bacteria cell wall derived agentssuch as peptidoglycan and other components derived from gram-positivebacteria with a high content of peptidoglycan. Exemplary gram-positivebacteria include Lactobacillus acidophilus, Bifedobact thermophilum,Bifedobat longhum, Streptococcus faecium, Bacillus pumilus, Bacillussubtilis, Bacillus licheniformis, Lactobacillus acidophilus,Lactobacillus casei, Enterococcus faecium, Bifidobacterium bifidium,Propionibacterium acidipropionici, Propionibacteriium freudenreichii,and Bifidobacterium pseudolongum.

The amount of formulated yeast can and will vary depending on theformulated yeast, the type of non-human animal that will be administeredthe feed composition, the body weight, sex, and medical condition of thenon-human animal among other variables, and can be determinedexperimentally. The concentration of formulated yeast in the feedcomposition may range from about 0.01 lb/ton of feed to about 1 lb/tonof feed, from about 0.1 lb/ton of feed to about 2 lb/ton of feed, fromabout 0.5 lb/ton of feed to about 1 lb/ton of feed, or from about 0.1lb/ton of feed to about 1 lb/ton of feed. Preferably, the concentrationof capsicum product in the feed composition ranges from about 0.3 lb/tonof feed to about 0.7 lb/ton of feed.

(c) Capsicum Product

The term “capsicum product” as used herein refers to any product derivedfrom the fruit of a plant in the Capsicum genus. Capsicum is a genus ofplants from the nightshade family (Solanaceae). The fruit of Capsicumplant have a variety of names. They are commonly called chili pepper,hot pepper, red or green pepper. Herein, they are referred to aspeppers. Some of these plants are used as spices, vegetables, or drugs.

The fruit of Capsicum plants generally comprise capsaicinoids.Capsaicinoids are the name given to the class of compounds found presentin members of the capsicum family of plants. Capsaicinoids arevanilloids, the heaviest of this class of compound in nature. Othervanilloids include vanillin, present in vanilla and the wood used to agewine, eugenol, present in bay leaves, allspice, and cloves, andzingerone, giving ginger and mustard their distinct flavors. The mostcommon capsaicinoid is N-Vanillyl-8-methyl-6-(E)-noneamide (capsaicin).Nearly as common is Dihydrocapsaicin. Other capsaicinoids arenordihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin. These andother capsaicinoids occur in varying ratios from plant to plant.

A capsicum product may be any product comprising one or more ofcapsaicin, dihydrocapsaicin, capsaicinoids, capsicum oleoresin,vanilloids, macerated hot peppers, ground hot peppers, extract from hotpeppers, other capsaicin- or capsicum-containing plant materials, anencapsulated or coated capsicum product, and combinations of anythereof.

A capsicum product may comprise one or more capsaicinoid. A capsaicinoidmay be synthetic capsaicinoids, a capsaicinoid extract from the fruitsof members of the capsicum family, or combinations thereof. Preferably,a capsaicinoid is a capsicum oleoresin. Capsicum oleoresin is aconcentrate of capsaicinoids that generally contains about 0.1% to 50%by weight of natural or synthetic capsaicinoids. Capsaicinoids,especially when concentrated in the form of capsicum oleoresin, aretoxic, requiring strict handling conditions not available for use byanimal feed manufacturers. As such, a capsaicinoid may be formulated forsafe handling. Methods of formulating capsaicinoids for safe handlingare known in the art. For instance, capsaicinoids may be in the form ofcapsules, microencapsulated formulations, caplets, and granules.

Preferably, a capsicum product comprises ground peppers. Ground peppersmay be the ground fruit of any Capsicum plant. Preferably, groundpeppers are the ground fruit of a hot pepper. Ground peppers may furtherbe formulated for safe and easy handling by animal feed manufacturers.For instance, ground pepper may be pelleted or encapsulated. A preferredcapsicum product is encapsulated ground pepper. More preferably, acapsicum product is encapsulated ground pepper comprising about 30% toabout 70% ground peppers, preferably about 40% to about 60% groundpeppers, and more preferably about 45% to about 55% ground peppers.Encapsulated ground peppers and methods of encapsulating ground peppersare known in the art.

Generally, the amount of capsicum product in a feed additive compositioncan and will vary depending on the capsicum product, the type ofnon-human animal that will be administered the feed additivecomposition, the body weight, sex, and medical condition of thenon-human animal that will be administered the feed additivecomposition.

Generally, a capsicum product comprises about 0.05% to about 1%capsaicinoids, preferably about 0.1% to about 0.7% capsaicinoids, andmore preferably about 0.4% to about 0.6% capsaicinoids. Irrespective ofthe capsaicinoid concentration in a capsicum product, a feed additivecomposition comprises an amount of capsicum product to provide fromabout 0.05% to about 5%, preferably from about 0.1% to about 2%capsaicinoids in the feed additive composition.

The amount of capsicum product in a feed composition can and will varydepending on the capsicum product, the type of non-human animal thatwill be administered the feed composition, the body weight, sex, andmedical condition of the non-human animal among other variables, and canbe determined experimentally. The concentration of capsicum product inthe feed composition may range from about 0.01 lb/ton of feed to about 1lb/ton of feed, from about 0.1 lb/ton of feed to about 2 lb/ton of feed,from about 0.5 lb/ton of feed to about 1 lb/ton of feed, or from fromabout 0.1 lb/ton of feed to about 1 lb/ton of feed. Preferably, theconcentration of capsicum product in the feed composition ranges fromabout 0.1 lb/ton of feed to about 0.5 lb/ton of feed.

(d) Antimicrobial Clay

As used herein, the term “antimicrobial clay” may be any clay havingantimicrobial properties. An antimicrobial clay may be capable ofcontrolling any one or more of bacteria, viruses, protozoans such asCryptosporidium spp. and giardia, and fungi such as mold and mildew. Asused herein, the term “antimicrobial” is used to indicate thatantimicrobial clay may either kill microbes, and therefore be“microbicidal,” or prevents microbes from growing and reproducing whilenot necessarily killing them otherwise, and therefore be “biostatic.”Methods of determining if an agent, including clay, has antimicrobialproperties are known in the art, and generally comprise contactingmicrobes with the agent in vivo or in vitro, and determining the effectof the agent on growth of the microbe.

Antimicrobial clays suitable for an animal feed composition are known inthe art, and may be as disclosed in U.S. application Ser. No.15/266,570. Preferably, an antimicrobial clay suitable for a feedcomposition of the disclosure comprises a naturally mined antimicrobialclay. Non-limiting examples of naturally-mined antimicrobial claysinclude clays supplied by Oregon Mineral Technologies (OMT; Grants Pass,Oreg.), also known as blue clay. The source of the blue clay is an openpit mine in hydrothermally altered, pyroclastic material in the CascadeMountains. Also preferred is a natural red clay mined in the CascadeMountain region of Oregon, more specifically a red clay mined in thecrater lake region of the Cascade Mountains of Oregon.

The amount of antimicrobial clay in a feed composition can and will varydepending on the antimicrobial clay, the type of non-human animal thatwill be administered the feed composition, the body weight, sex, andmedical condition of the non-human animal among other variables, and canbe determined experimentally. The concentration of antimicrobial clay inthe feed composition may range from about 0.1 lb/ton of feed to about100 lb/ton of feed, from about 0.1 lb/ton of feed to about 10 lb/ton offeed, from about 1 lb/ton of feed to about 50 lb/ton of feed, or fromfrom about 1 lb/ton of feed to about 10 lb/ton of feed. Preferably, theconcentration of antimicrobial clay in the feed composition ranges fromabout 2 lb/ton of feed to about 6 lb/ton of feed.

(e) Basal Animal Diet

A basal animal diet suitable for a feed composition of the disclosurecan and will vary depending on the intended animal, the weight of theanimal, and the stage of development of the animal among othervariables.

The terms “feed”, “food”, and “feed formulation” are used hereininterchangeably and may refer to any feed composition normally fed to ananimal. Basal animal diets normally fed to an animal are known in theart. A basal animal diet may include one or more components of an animalfeed. Non-limiting examples of feed matter or animal feed matter mayinclude, without limitation: corn or a component of corn, such as, forexample, corn meal, corn fiber, corn hulls, corn DDGS (distiller's driedgrain with solubles), silage, ground corn, corn germ, corn gluten, cornoil, or any other portion of a corn plant; soy or a component of soy,such as, for example, soy oil, soy meal, soy hulls, soy silage, groundsoy, or any other portion of a soy plant; wheat or any component ofwheat, such as, for example, wheat meal, wheat fiber, wheat hulls, wheatchaff, ground wheat, wheat germ, or any other portion of a wheat plant;rice or any component of rice, such as, for example, rice meal, ricefiber, rice hulls, rice chaff, ground rice, rice germ, or any otherportion of a rice plant; canola, such as, for example, canola oil,canola meal, canola protein, canola hulls, ground canola, or any otherportion of a canola plant; sunflower or a component of a sunflowerplant; sorghum or a component of a sorghum plant; sugar beet or acomponent of a sugar beet plant; cane sugar or a component of asugarcane plant; barley or a component of a barley plant; palm oil, palmkernel or a component of a palm plant; glycerol; corn steep liquor; awaste stream from an agricultural processing facility; lecithin; rumenprotected fats; molasses; soy molasses; flax; peanuts; peas; oats;grasses, such as orchard grass and fescue; fish meal, meat & bone meal;feather meal; and poultry byproduct meal; and alfalfa and/or clover usedfor silage or hay, and various combinations of any of the feedingredients set forth herein, or other feed ingredients generally knownin the art. As it will be recognized in the art, a basal animal diet mayfurther be supplemented with amino acids, vitamins, minerals, and otherfeed additives such as other types of enzymes, organic acids, essentialoils, probiotics, prebiotics, antioxidants, pigments, anti-cakingagents, and the like, as described further below. A basal animal dietmay be formulated for administration to any animal subject. Animalsubjects may be as described below.

The basal animal diets may optionally comprise at least one additionalnutritive and/or pharmaceutical agent. For instance, the at least oneadditional nutritive and/or pharmaceutical agent may be selected fromthe group consisting of vitamin, mineral, amino acid, antioxidant,probiotic, essential fatty acid, and pharmaceutically acceptableexcipient. The compositions may include one additional nutritive and/orpharmaceutical component or a combination of any of the foregoingadditional components in varying amounts. Suitable examples of eachadditional component are detailed below.

(i) Vitamins

Optionally, the animal feed formulation may include one or morevitamins. Suitable vitamins for use in the dietary supplement includevitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin,niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine,pantothenic acid, and biotin. The form of the vitamin may include saltsof the vitamin, derivatives of the vitamin, compounds having the same orsimilar activity of a vitamin, and metabolites of a vitamin.

The animal feed formulation may include one or more forms of aneffective amount of any of the vitamins described herein or otherwiseknown in the art. Exemplary vitamins include vitamin K, vitamin D,vitamin C, and biotin. An “effective amount” of a vitamin typicallyquantifies an amount at least about 10% of the United States RecommendedDaily Allowance (“RDA”) of that particular vitamin for a subject. It iscontemplated, however, that amounts of certain vitamins exceeding theRDA may be beneficial for certain animals. For example, the amount of agiven vitamin may exceed the applicable RDA by 100%, 200%, 300%, 400%,500% or more.

(ii) Minerals

Generally, the animal feed formulation may include one or more mineralsor mineral sources. Non-limiting examples of minerals include, withoutlimitation, calcium, iron, chromium, copper, iodine, zinc, magnesium,manganese, molybdenum, phosphorus, potassium, and selenium. Suitableforms of any of the foregoing minerals include soluble mineral salts,slightly soluble mineral salts, insoluble mineral salts, chelatedminerals, mineral complexes, non-reactive minerals such as carbonylminerals, and reduced minerals, and combinations thereof.

Generally speaking, the animal feed formulation may include one or moreforms of an effective amount of any of the minerals described herein orotherwise known in the art. An “effective amount” of a mineral typicallyquantifies an amount at least about 10% of the United States RecommendedDaily Allowance (“RDA”) of that particular mineral for a subject. It iscontemplated, however, that amounts of certain minerals exceeding theRDA may be beneficial for certain subjects. For example, the amount of agiven mineral may exceed the applicable RDA by 100%, 200%, 300%, 400%,500% or more. Typically, the amount of mineral included in the dietarysupplement may range from about 1 mg to about 1500 mg, about 5 mg toabout 500 mg, or from about 50 mg to about 500 mg per dosage.

(iii) Essential Fatty Acids

Optionally, the animal feed formulation may include a source of anessential fatty acid. The essential fatty acid may be isolated or it maybe an oil source or fat source that contains an essential fatty acid. Inone embodiment, the essential fatty acid may be a polyunsaturated fattyacid (PUFA), which has at least two carbon-carbon double bonds generallyin the cis-configuration. The PUFA may be a long chain fatty acid havingat least 18 carbons atoms. The PUFA may be an omega-3 fatty acid inwhich the first double bond occurs in the third carbon-carbon bond fromthe methyl end of the carbon chain (i.e., opposite the carboxyl acidgroup). Examples of omega-3 fatty acids include alpha-linolenic acid(18:3, ALA), stearidonic acid (18:4), eicosatetraenoic acid (20:4),eicosapentaenoic acid (20:5; EPA), docosatetraenoic acid (22:4), n-3docosapentaenoic acid (22:5; n-3DPA), and docosahexaenoic acid (22:6;DHA). The PUFA may also be an omega-5 fatty acid, in which the firstdouble bond occurs in the fifth carbon-carbon bond from the methyl end.Exemplary omega-5 fatty acids include myristoleic acid (14:1),myristoleic acid esters, and cetyl myristoleate. The PUFA may also be anomega-6 fatty acid, in which the first double bond occurs in the sixthcarbon-carbon bond from the methyl end. Examples of omega-6 fatty acidsinclude linoleic acid (18:2), gamma-linolenic acid (18:3), eicosadienoicacid (20:2), dihomo-gamma-linolenic acid (20:3), arachidonic acid(20:4), docosadienoic acid (22:2), adrenic acid (22:4), and n-6docosapentaenoic acid (22:5). The fatty acid may also be an omega-9fatty acid, such as oleic acid (18:1), eicosenoic acid (20:1), mead acid(20:3), erucic acid (22:1), and nervonic acid (24:1).

In another embodiment, the essential fatty acid source may be aseafood-derived oil. The seafood may be a vertebrate fish or a marineorganism, such that the oil may be fish oil or marine oil. The longchain (20C, 22C) omega-3 and omega-6 fatty acids are found in seafood.The ratio of omega-3 to omega-6 fatty acids in seafood ranges from about8:1 to 20:1. Seafood from which oil rich in omega-3 fatty acids may bederived include, but are not limited to, abalone scallops, albacoretuna, anchovies, catfish, clams, cod, gem fish, herring, lake trout,mackerel, menhaden, orange roughy, salmon, sardines, sea mullet, seaperch, shark, shrimp, squid, trout, and tuna.

In yet another embodiment, the essential fatty acid source may be aplant-derived oil. Plant and vegetable oils are rich in omega-6 fattyacids. Some plant-derived oils, such as flaxseed oil, are especiallyrich in omega-3 fatty acids. Plant or vegetable oils are generallyextracted from the seeds of a plant, but may also be extracted fromother parts of the plant. Plant or vegetable oils that are commonly usedfor cooking or flavoring include, but are not limited to, acai oil,almond oil, amaranth oil, apricot seed oil, argan oil, avocado seed oil,babassu oil, ben oil, blackcurrant seed oil, Borneo tallow nut oil,borage seed oil, buffalo gourd oil, canola oil, carob pod oil, cashewoil, castor oil, coconut oil, coriander seed oil, corn oil, cottonseedoil, evening primrose oil, false flax oil, flax seed oil, grapeseed oil,hazelnut oil, hemp seed oil, kapok seed oil, lallemantia oil, linseedoil, macadamia oil, meadowfoam seed oil, mustard seed oil, okra seedoil, olive oil, palm oil, palm kernel oil, peanut oil, pecan oil, pequioil, perilla seed oil, pine nut oil, pistachio oil, poppy seed oil,prune kernel oil, pumpkin seed oil, quinoa oil, ramtil oil, rice branoil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil,thistle oil, walnut oil, or wheat germ oil. The plant-derived oil mayalso be hydrogenated or partially hydrogenated.

In still a further embodiment, the essential fatty acid source may be analgae-derived oil. Commercially available algae-derived oils includethose from Crypthecodinium cohnii and Schizochytrium sp. Other suitablespecies of algae, from which oil is extracted, include Aphanizomenonflos-aquae, Bacilliarophy sp., Botryococcus braunii, Chlorophyceae sp.,Dunaliella tertiolecta, Euglena gracilis, Isochrysis galbana,Nannochloropsis salina, Nannochloris sp., Neochloris oleoabundans,Phaeodactylum tricornutum, Pleurochrysis carterae, Prymnesium parvum,Scenedesmus dimorphus, Spirulina sp., and Tetraselmis chui.

(iv) Amino Acids

The animal feed formulation may optionally include from one to severalamino acids. Suitable amino acids include alanine, arginine, asparagine,aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine,isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine,threonine, tryptophan, tyrosine, and valine or their hydroxy analogs. Incertain embodiments, the amino acid will be selected from the essentialamino acids. An essential amino acid is generally described as one thatcannot be synthesized de novo by the organism, and therefore, must beprovided in the diet. By way of non-limiting example, the essentialamino acids for humans include: L-histidine, L-isoleucine, L-leucine,L-lysine, L-methionine, L-phenylalanine, L-valine and L-threonine.

(v) Antioxidants

The animal feed formulation may include one or more suitableantioxidants. As will be appreciated by a skilled artisan, thesuitability of a given antioxidant will vary depending upon the speciesto which the dietary supplement will be administered. Non-limitingexamples of antioxidants include ascorbic acid and its salts, ascorbylpalmitate, ascorbyl stearate, anoxomer, N-acetylcysteine, benzylisothiocyanate, o-, m- or p-amino benzoic acid (o is anthranilic acid, pis PABA), butylated hydroxyanisole (BHA), butylated hydroxytoluene(BHT), caffeic acid, canthaxantin, alpha-carotene, beta-carotene,beta-caraotene, beta-apo-carotenoic acid, carnosol, carvacrol,catechins, cetyl gallate, chlorogenic acid, citric acid and its salts,p-coumaric acid, curcurin, 3,4-dihydroxybenzoic acid,N,N′-diphenyl-p-phenylenediamine (DPPD), dilauryl thiodipropionate,distearyl thiodipropionate, 2,6-di-tert-butylphenol, dodecyl gallate,edetic acid, ellagic acid, erythorbic acid, sodium erythorbate,esculetin, esculin, 6-ethoxy-1,2-dihydro-2,2,4-trimethylquinoline, ethylgallate, ethyl maltol, ethylenediaminetetraacetic acid (EDTA), eugenol,ferulic acid, flavonoids, flavones (e.g., apigenin, chrysin, luteolin),flavonols (e.g., datiscetin, myricetin, daemfero), flavanones, fraxetin,fumaric acid, gallic acid, gentian extract, gluconic acid, glycine, gumguaiacum, hesperetin, alpha-hydroxybenzyl phosphinic acid,hydroxycinammic acid, hydroxyglutaric acid, hydroquinone,N-hydroxysuccinic acid, hydroxytryrosol, hydroxyurea, lactic acid andits salts, lecithin, lecithin citrate; R-alpha-lipoic acid, lutein,lycopene, malic acid, maltol, 5-methoxy tryptamine, methyl gallate,monoglyceride citrate; monoisopropyl citrate; morin,beta-naphthoflavone, nordihydroguaiaretic acid (NDGA), octyl gallate,oxalic acid, palmityl citrate, phenothiazine, phosphatidylcholine,phosphoric acid, phosphates, phytic acid, phytylubichromel, propylgallate, polyphosphates, quercetin, trans-resveratrol, rosmarinic acid,sesamol, silymarin, sinapic acid, succinic acid, stearyl citrate,syringic acid, tartaric acid, thymol, tocopherols (i.e., alpha-, beta-,gamma- and delta-tocopherol), tocotrienols (i.e., alpha-, beta-, gamma-and delta-tocotrienols), tyrosol, vanilic acid,2,6-di-tert-butyl-4-hydroxymethylphenol (i.e., lonox 100),2,4-(tris-3′,5′-bi-tert-butyl-4′-hydroxybenzyl)-mesitylene (i.e., lonox330), 2,4,5-trihydroxybutyrophenone, ubiquinone, tertiary butylhydroquinone (TBHQ), thiodipropionic acid, trihydroxy butyrophenone,tryptamine, tyramine, uric acid, vitamin K and derivates, vitamin Q10,zeaxanthin, or combinations thereof.

Natural antioxidants that may be included in the dietary supplementinclude, but are not limited to, apple peel extract, blueberry extract,carrot juice powder, clove extract, coffee berry, coffee bean extract,cranberry extract, eucalyptus extract, ginger powder, grape seedextract, green tea, olive leaf, parsley extract, peppermint, pimentoextract, pomace, pomegranate extract, rice bran extract, rosehips,rosemary extract, sage extract, tart cherry extract, tomato extract,turmeric, and wheat germ oil.

(vi) Anti-Inflammatory Agents

The animal feed formulation may optionally include at least oneanti-inflammatory agent. In one embodiment, the anti-inflammatory agentmay be a synthetic non-steroidal anti-inflammatory drug (NSAID) such asacetylsalicylic acid, dichlophenac, indomethacin, oxamethacin,ibuprofen, indoprofen, naproxen, ketoprofen, mefamanic acid, metamizole,piroxicam, and celecoxib. In an alternate embodiment, theanti-inflammatory agent may be a prohormone that modulates inflammatoryprocesses. Suitable prohormones having this property include prohormoneconvertase 1, proopiomelanocortin, prohormone B-type natriureticpeptide, SMR1 prohormone, and the like. In another embodiment, theanti-inflammatory agent may be an enzyme having anti-inflammatoryeffects. Examples of anti-inflammatory enzymes include bromelain,papain, serrapeptidase, and proteolytic enzymes such as pancreatin (amixture of trypsin, amylase and lipase).

In still another embodiment, the anti-inflammatory agent may be apeptide with anti-inflammatory effects. For example, the peptide may bean inhibitor of phospholipase A2, such as antiflammin-1, a peptide thatcorresponds to amino acid residues 246-254 of lipocortin; antiflammin-2,a peptide that corresponds to amino acid residues 39-47 of uteroglobin;S7 peptide, which inhibits the interaction between interleukin 6 andinterleukin 6 receptor; RP1, a prenyl protein inhibitor; and similarpeptides. Alternatively, the anti-inflammatory peptide may becortistatin, a cyclic neuropeptide related to somatostatin, or peptidesthat correspond to an N-terminal fragment of SV-IV protein, a conservedregion of E-, L-, and P-selectins, and the like. Other suitableanti-inflammatory preparations include collagen hydrolysates and milkmicronutrient concentrates (e.g., MicroLactin® available from StolleMilk Biologics, Inc., Cincinnati, Ohio), as well as milk proteinhydrolysates, casein hydrolysates, whey protein hydrolysates, and plantprotein hydrolysates.

In a further embodiment, the anti-inflammatory agent may be a probioticthat has been shown to modulate inflammation. Suitable immunomodulatoryprobiotics include lactic acid bacteria such as acidophilli,lactobacilli, and bifidophilli. In yet another embodiment, theanti-inflammatory agent may be a plant extract having anti-inflammatoryproperties. Non-limiting examples of suitable plant extracts withanti-inflammatory benefits include blueberries, boswella, black catechuand Chinese skullcap, celery seed, chamomile, cherries, devils claw,eucalyptus, evening primrose, ginger, hawthorne berries, horsetail,Kalopanax pictus bark, licorice root, turmeric, white wallow, willowbark, and yucca.

(vii) Herbals

The animal feed formulation may optionally include at least one herb orherbal derivative. Suitable herbals and herbal derivatives, as usedherein, refer to herbal extracts, and substances derived from plants andplant parts, such as leaves, flowers, and roots, without limitation.Non-limiting exemplary herbals and herbal derivatives include agrimony,alfalfa, aloe vera, amaranth, angelica, anise, barberry, basil,bayberry, bee pollen, birch, bistort, blackberry, black cohosh, blackwalnut, blessed thistle, blue cohosh, blue vervain, boneset, borage,buchu, buckthorn, bugleweed, burdock, capsicum, cayenne, caraway,cascara sagrada, catnip, celery, centaury, chamomile, chaparral,chickweed, chicory, chinchona, cloves, coltsfoot, comfrey, cornsilk,couch grass, cramp bark, culver's root, cyani, cornflower, damiana,dandelion, devils claw, dong quai, echinacea, elecampane, ephedra,eucalyptus, evening primrose, eyebright, false unicorn, fennel,fenugreek, figwort, flaxseed, garlic, gentian, ginger, ginseng, goldenseal, gotu kola, gum weed, hawthorn, hops, horehound, horseradish,horsetail, hoshouwu, hydrangea, hyssop, iceland moss, irish moss,jojoba, juniper, kelp, lady's slipper, lemon grass, licorice, lobelia,mandrake, marigold, marjoram, marshmallow, mistletoe, mullein, mustard,myrrh, nettle, oatstraw, oregon grape, papaya, parsley, passion flower,peach, pennyroyal, peppermint, periwinkle, plantain, pleurisy root,pokeweed, prickly ash, psyllium, quassia, queen of the meadow, redclover, red raspberry, redmond clay, rhubarb, rose hips, rosemary, rue,safflower, saffron, sage, St. John's wort, sarsaparilla, sassafras, sawpalmetto, skullcap, senega, senna, shepherd's purse, slippery elm,spearmint, spikenard, squawvine, stillingia, strawberry, taheebo, thyme,uva ursi, valerian, violet, watercress, white oak bark, white pine bark,wild cherry, wild lettuce, wild yam, willow, wintergreen, witch hazel,wood betony, wormwood, yarrow, yellow dock, yerba santa, yucca andcombinations thereof.

(viii) Pigments

The animal feed formulation may optionally include at least one pigment.Suitable non-limiting pigments include actinioerythrin, alizarin,alloxanthin, β-apo-2′-carotenal, apo-2-lycopenal, apo-6′-lycopenal,astacein, astaxanthin, azafrinaldehyde, aacterioruberin, aixin,α-carotine, β-carotine, γ-carotine, β-carotenone, canthaxanthin,capsanthin, capsorubin, citranaxanthin, citroxanthin, crocetin,crocetinsemialdehyde, crocin, crustaxanthin, cryptocapsin,α-cryptoxanthin, β-cryptoxanthin, cryptomonaxanthin, cynthiaxanthin,decaprenoxanthin, dehydroadonirubin, diadinoxanthin,1,4-diamino-2,3-dihydroanthraquinone, 1,4-dihydroxyanthraquinone,2,2′-diketospirilloxanthin, eschscholtzxanthin, eschscholtzxanthone,flexixanthin, foliachrome, fucoxanthin, gazaniaxanthin,hexahydrolycopene, hopkinsiaxanthin, hydroxyspheriodenone,isofucoxanthin, loroxanthin, lutein, luteoxanthin, lycopene,lycopersene, lycoxanthin, morindone, mutatoxanthin, neochrome,neoxanthin, nonaprenoxanthin, OH-Chlorobactene, okenone, oscillaxanthin,paracentrone, pectenolone, pectenoxanthin, peridinin, phleixanthophyll,phoeniconone, phoenicopterone, phoenicoxanthin, physalien, phytofluene,pyrrhoxanthininol, quinones, rhodopin, rhodopinal, rhodopinol,rhodovibrin, rhodoxanthin, rubixanthone, saproxanthin,semi-α-carotenone, semi-β-carotenone, sintaxanthin, siphonaxanthin,siphonein, spheroidene, tangeraxanthin, torularhodin, torularhodinmethyl ester, torularhodinaldehyde, torulene,1,2,4-trihydroxyanthraquinone, triphasiaxanthin, trollichrome,vaucheriaxanthin, violaxanthin, wamingone, xanthin, zeaxanthin,α-zeacarotene, or combinations thereof.

(ix) Pharmaceutical Acceptable Agents

The animal feed formulation may optionally include at least onepharmaceutical acceptable agent. Suitable non-limiting pharmaceuticallyacceptable agents include an acid/alkaline-labile drug, a pH dependentdrug, or a drug that is a weak acid or a weak base. Examples ofacid-labile drugs include statins (e.g., pravastatin, fluvastatin andatorvastatin), antiobiotics (e.g., penicillin G, ampicillin,streptomycin, erythromycin, clarithromycin and azithromycin), nucleosideanalogs (e.g., dideoxyinosine (ddI or didanosine), dideoxyadenosine(ddA), dideoxycytosine (ddC), salicylates (e.g., aspirin), digoxin,bupropion, pancreatin, midazolam, and methadone. Drugs that are onlysoluble at acid pH include nifedipine, emonapride, nicardipine,amosulalol, noscapine, propafenone, quinine, dipyridamole, josamycin,dilevalol, labetalol, enisoprost, and metronidazole. Drugs that are weakacids include phenobarbital, phenytoin, zidovudine (AZT), salicylates(e.g., aspirin), propionic acid compounds (e.g., ibuprofen), indolederivatives (e.g., indomethacin), fenamate compounds (e.g., meclofenamicacid), pyrrolealkanoic acid compounds (e.g., tolmetin), cephalosporins(e.g., cephalothin, cephalaxin, cefazolin, cephradine, cephapirin,cefamandole, and cefoxitin), 6-fluoroquinolones, and prostaglandins.Drugs that are weak bases include adrenergic agents (e.g., ephedrine,desoxyephedrine, phenylephrine, epinephrine, salbutamol, andterbutaline), cholinergic agents (e.g., physostigmine and neostigmine),antispasmodic agents (e.g., atropine, methantheline, and papaverine),curariform agents (e.g., chlorisondamine), tranquilizers and musclerelaxants (e.g., fluphenazine, thioridazine, trifluoperazine,chlorpromazine, and triflupromazine), antidepressants (e.g.,amitriptyline and nortriptyline), antihistamines (e.g., diphenhydramine,chlorpheniramine, dimenhydrinate, tripelennamine, perphenazine,chlorprophenazine, and chlorprophenpyridamine), cardioactive agents(e.g., verapamil, diltiazem, gallapomil, cinnarizine, propranolol,metoprolol and nadolol), antimalarials (e.g., chloroquine), analgesics(e.g., propoxyphene and meperidine), antifungal agents (e.g.,ketoconazole and itraconazole), antimicrobial agents (e.g., cefpodoxime,proxetil, and enoxacin), caffeine, theophylline, and morphine. Inanother embodiment, the drug may be a biphosphonate or another drug usedto treat osteoporosis. Non-limiting examples of a biphosphonate includealendronate, ibandronate, risedronate, zoledronate, pamidronate,neridronate, olpadronate, etidronate, clodronate, and tiludronate. Othersuitable drugs include estrogen, selective estrogen receptor modulators(SERMs), and parathyroid hormone (PTH) drugs. In yet another embodiment,the drug may be an antibacterial agent. Suitable antibiotics includeaminoglycosides (e.g., amikacin, gentamicin, kanamycin, neomycin,netilmicin, streptomycin, and tobramycin), carbecephems (e.g.,loracarbef), a carbapenem (e.g., certapenem, imipenem, and meropenem),cephalosporins (e.g., cefadroxil cefazolin, cephalexin, cefaclor,cefamandole, cephalexin, cefoxitin, cefprozil, cefuroxime, cefixime,cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime,ceftazidime, ceftibuten, ceftizoxime, and ceftriaxone), macrolides(e.g., azithromycin, clarithromycin, dirithromycin, erythromycin, andtroleandomycin), monobactam, penicillins (e.g., amoxicillin, ampicillin,carbenicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin,penicillin G, penicillin V, piperacillin, and ticarcillin), polypeptides(e.g., bacitracin, colistin, and polymyxin B), quinolones (e.g.,ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin,moxifloxacin, norfloxacin, ofloxacin, and trovafloxacin), sulfonamides(e.g., mafenide, sulfacetamide, sulfamethizole, sulfasalazine,sulfisoxazole, and trimethoprim-sulfamethoxazole), and tetracyclines(e.g., demeclocycline, doxycycline, minocycline, and oxytetracycline).In an alternate embodiment, the drug may be an antiviral proteaseinhibitor (e.g., amprenavir, fosamprenavir, indinavir,lopinavir/ritonavir, ritonavir, saquinavir, and nelfinavir). In stillanother embodiment, the drug may be a cardiovascular drug. Examples ofsuitable cardiovascular agents include cardiotonic agents (e.g.,digitalis (digoxin), ubidecarenone, and dopamine), vasodilating agents(e.g., nitroglycerin, captopril, dihydralazine, diltiazem, andisosorbide dinitrate), antihypertensive agents (e.g., alpha-methyldopa,chlortalidone, reserpine, syrosingopine, rescinnamine, prazosin,phentolamine, felodipine, propanolol, pindolol, labetalol, clonidine,captopril, enalapril, and lisonopril), beta blockers (e.g., levobunolol,pindolol, timolol maleate, bisoprolol, carvedilol, and butoxamine),alpha blockers (e.g., doxazosin, prazosin, phenoxybenzamine,phentolamine, tamsulosin, alfuzosin, and terazosin), calcium channelblockers (e.g., amlodipine, felodipine, nicardipine, nifedipine,nimodipine, nisoldipine, nitrendipine, lacidipine, lercanidipine,verapamil, gallopamil, and diltiazem), monensin, avilamycin,salinomycin, narasin, diclaserol, tylosin, bacitracin, bacitracin zinc,and anticlot agents (e.g., dipyrimadole).

(x) Excipients

A variety of commonly used excipients in animal feed formulation may beselected on the basis of compatibility with the active ingredients.Non-limiting examples of suitable excipients include an agent selectedfrom the group consisting of non-effervescent disintegrants, a coloringagent, a flavor-modifying agent, an oral dispersing agent, a stabilizer,a preservative, a diluent, a compaction agent, a lubricant, a filler, abinder, taste masking agents, an effervescent disintegration agent, andcombinations of any of these agents.

In one embodiment, the excipient is a binder. Suitable binders includestarches, pregelatinized starches, gelatin, polyvinylpyrolidone,cellulose, methylcellulose, sodium carboxymethylcellulose,ethylcellulose, polyacrylamides, polyvinyloxoazolidone,polyvinylalcohols, C12-C18 fatty acid alcohol, polyethylene glycol,polyols, saccharides, oligosaccharides, polypeptides, oligopeptides, andcombinations thereof. The polypeptide may be any arrangement of aminoacids ranging from about 100 to about 300,000 daltons.

In another embodiment, the excipient may be a filler. Suitable fillersinclude carbohydrates, inorganic compounds, and polyvinylpirrolydone. Byway of non-limiting example, the filler may be calcium sulfate, both di-and tri-basic, starch, calcium carbonate, magnesium carbonate,microcrystalline cellulose, dibasic calcium phosphate, magnesiumcarbonate, magnesium oxide, calcium silicate, talc, modified starches,lactose, sucrose, mannitol, and sorbitol.

The excipient may comprise a non-effervescent disintegrant. Suitableexamples of non-effervescent disintegrants include starches such as cornstarch, potato starch, pregelatinized and modified starches thereof,sweeteners, clays, such as bentonite, micro-crystalline cellulose,alginates, sodium starch glycolate, gums such as agar, guar, locustbean, karaya, pecitin, and tragacanth.

In another embodiment, the excipient may be an effervescentdisintegrant. By way of non-limiting example, suitable effervescentdisintegrants include sodium bicarbonate in combination with citric acidand sodium bicarbonate in combination with tartaric acid.

The excipient may comprise a preservative. Suitable examples ofpreservatives include antioxidants, such as a-tocopherol or ascorbate,and antimicrobials, such as parabens, chlorobutanol or phenol.

In another embodiment, the excipient may include a diluent. Diluentssuitable for use include pharmaceutically acceptable saccharides such assucrose, dextrose, lactose, microcrystalline cellulose, fructose,xylitol, and sorbitol; polyhydric alcohols; a starch; pre-manufactureddirect compression diluents; and mixtures of any of the foregoing.

The excipient may include flavors. Flavors incorporated into the outerlayer may be chosen from synthetic flavor oils and flavoring aromaticsand/or natural oils, extracts from plants, leaves, flowers, fruits, andcombinations thereof. By way of example, these may include cinnamonoils, oil of wintergreen, peppermint oils, clover oil, hay oil, aniseoil, eucalyptus, vanilla, citrus oil, such as lemon oil, orange oil,grape and grapefruit oil, fruit essences including apple, peach, pear,strawberry, raspberry, cherry, plum, pineapple, and apricot.

In another embodiment, the excipient may include a sweetener. By way ofnon-limiting example, the sweetener may be selected from glucose (cornsyrup), dextrose, invert sugar, fructose, and mixtures thereof (when notused as a carrier); saccharin and its various salts such as the sodiumsalt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds,glycyrrhizin; Stevia Rebaudiana (Stevioside); chloro derivatives ofsucrose such as sucralose; sugar alcohols such as sorbitol, mannitol,sylitol, and the like.

In another embodiment, the excipient may be a lubricant. Suitablenon-limiting examples of lubricants include magnesium stearate, calciumstearate, zinc stearate, hydrogenated vegetable oils, sterotex,polyoxyethylene monostearate, talc, polyethyleneglycol, sodium benzoate,sodium lauryl sulfate, magnesium lauryl sulfate, and light mineral oil.

The excipient may be a dispersion enhancer. Suitable dispersants mayinclude starch, alginic acid, polyvinylpyrrolidones, guar gum, kaolin,bentonite, purified wood cellulose, sodium starch glycolate,isoamorphous silicate, and microcrystalline cellulose as high HLBemulsifier surfactants.

Depending upon the embodiment, it may be desirable to provide a coloringagent in the outer layer. Suitable color additives include food, drugand cosmetic colors (FD&C), drug and cosmetic colors (D&C), or externaldrug and cosmetic colors (Ext. D&C). These colors or dyes, along withtheir corresponding lakes, and certain natural and derived colorants,may be suitable for use in the present invention depending on theembodiment.

The excipient may include a taste-masking agent. Taste-masking materialsinclude, e.g., cellulose hydroxypropyl ethers (HPC) such as Klucel®,Nisswo HPC and PrimaFlo HP22; low-substituted hydroxypropyl ethers(L-HPC); cellulose hydroxypropyl methyl ethers (HPMC) such asSeppifilm-LC, Pharmacoat®, Metolose SR, Opadry YS, PrimaFlo, MP3295A,Benecel MP824, and Benecel MP843; methylcellulose polymers such asMethocel® and Metolose®; Ethylcelluloses (EC) and mixtures thereof suchas E461, Ethocel®, Aqualon®-EC, Surelease; Polyvinyl alcohol (PVA) suchas Opadry AMB; hydroxyethylcelluloses such as Natrosol®;carboxymethylcelluloses and salts of carboxymethylcelluloses (CMC) suchas Aualon®-CMC; polyvinyl alcohol and polyethylene glycol co-polymerssuch as Kollicoat IR®; monoglycerides (Myverol), triglycerides (KLX),polyethylene glycols, modified food starch, acrylic polymers andmixtures of acrylic polymers with cellulose ethers such as Eudragit®EPO, Eudragit® RD100, and Eudragit® E100; cellulose acetate phthalate;sepifilms such as mixtures of HPMC and stearic acid, cyclodextrins, andmixtures of these materials. In other embodiments, additionaltaste-masking materials contemplated are those described in U.S. Pat.Nos. 4,851,226; 5,075,114; and 5,876,759, each of which is herebyincorporated by reference in its entirety.

In various embodiments, the excipient may include a pH modifier. Incertain embodiments, the pH modifier may include sodium carbonate orsodium bicarbonate.

The amount and types of ingredients (i.e., metal chelate, chondroprotective agents, vitamin, mineral, amino acid, antioxidant, yeastculture, and essential fatty acid), and other excipients useful inanimal feed formulation, are described throughout the specification andthe examples.

(II) Feed Additive Composition

One aspect of the present disclosure encompasses feed additivecompositions for non-human animals comprising at least two of a directfed microbial (DFM), a formulated yeast, a capsicum product, and anantimicrobial clay. Other optional additives may be further included.The feed additive composition may be added to basal animal diet foradministration to the non-human animals. The feed additive compositionmay be formulated with basal animal diets to prepare the feedcompositions described in Section I.

In some embodiments, formulated yeast in a basal animal diet comprisesabout 80-100% yeast extract, more preferably about 99% yeast extract.The amount of formulated yeast in a feed additive composition can andwill vary depending on the formulated yeast, the type of non-humananimal that will be administered the feed additive compositioncomprising the formulated yeast, the body weight, sex, and medicalcondition of the non-human animal that will be administered the feedadditive composition. Generally, a feed additive composition comprisesabout 30% to about 70% formulated yeast, preferably about 40% to about60% formulated yeast, and more preferably about 45% to about 55%formulated yeast.

When a capsicum product comprises ground peppers, a feed additivecomposition comprises about 5% to about 25% ground peppers, preferablyabout 10% to about 20% ground peppers. Preferably, the ground peppersare from a capsicum product comprising encapsulated ground pepper in anamount ranging about 30% to about 70% ground peppers, preferably about40% to about 60% ground peppers, and more preferably about 45% to about55% ground peppers.

In various embodiments, a feed additive composition may be introduced toa basal animal diet by way of various methods, depending on whether thefeed additive composition is in a liquid or solid form. Non-limitingexamples of introducing the feed additive composition to a basal animaldiet may be formulating the feed additive composition into the basalanimal diet, top-dressing the solid composition of a basal animal diet,spraying a liquid feed additive composition onto a basal animal diet, orcombinations thereof. It will be recognized that, when the feed additiveis introduced to a basal animal diet, the amount of the feed additiveintroduced to a basal animal diet is sufficient to provide thetherapeutically effective amount of the combination of formulated yeastand capsicum product in the diet of the animal.

In addition to DFM and at least one of a formulated yeast, a capsicumproduct, and an antimicrobial clay, a feed additive composition mayfurther comprise at least one additional ingredient such as vitamins,minerals, amino acids, antioxidants, probiotics, essential fatty acids,and pharmaceutically acceptable excipients. Such ingredients may be asdescribed in Section I(e) above. In some embodiments, a feed additivecomposition further comprises calcium carbonate, rice hulls, mineraloil, and calcium stearate.

(III) Methods of Using

Another aspect of the disclosure encompasses methods of using a feedcomposition. The methods comprise administering the animal feedcomposition to non-human animals. Preferably, a feed composition isadministered to non-human animals orally. A feed composition may be asdescribed in Section (I).

The timing and duration of administration of a composition of theinvention to an animal can and will vary. The feed composition may beadministered throughout the period of feeding the animal. Alternatively,the feed composition may be administered at specific periods during thegrowth and development of the animal. For instance, the feed compositionmay be administered during periods of heightened susceptibility of theanimal to infection, such as during infancy. A composition may also beadministered after a microbial infection is detected and for theduration of the infection. A composition may also be administered atvarious intervals. For instance, a composition may be administereddaily, weekly, monthly, or over a number of months. In some embodiments,a composition is administered weekly. In other embodiments, acomposition is administered monthly. In preferred embodiments, acomposition is administered daily. As it will be recognized in the art,the duration of treatment can and will vary depending on the growth andhealth of the animal.

(a) Non-Human Animals

Non-human animals, in broad term, may be defined as any animal whichexhibits improved growth, improved health, improved intestinal health,and reduced microbial pathogen counts after administration of the feedadditive composition. In various embodiments, the non-human animal maybe a livestock mammal varying in age and health. Non-limiting example ofsuitable livestock mammals may be beef cattle, horses, dairy cattle,veal, pigs, goats, sheep, bison, llama, or alpaca. In other embodiments,the non-human animal may be an avian species varying in age and health.Non-limiting examples of suitable avian species or poultry may bechickens, including broilers, layers, and breeders, ducks, game hens,geese, pheasants, guinea fowl/hens, quail, turkeys, and ratites, such asemus and ostrich. In another embodiment, the non-human animal may be acompanion animal varying in age and health. Non-limiting examples ofcompanion animals may be a dog, a cat, a bird, a hamster, or a Guineepig. In a preferred embodiment, the non-human animal is selected from agroup comprising growing pigs, calves, foals, kids (goats), lambs, cria,chicks, poults, ducklings, puppies, kittens, or combinations thereof.Most preferred, the non-human animals are chickens varying in age andhealth.

(b) Improved Performance

In some embodiments, the methods comprise using the feed additive toimprove performance of the animal. “Improved performance”, as definedherein, refers to a positive change in size and/or maturation over aperiod of time in the non-human animal. In various embodiments, thenon-human animals may exhibit improved growth, including for example anincrease in body weight gain, feed intake, average daily weight gain(ADG), a decrease in the feed conversion rate (FCR), an increase in theaverage daily food intake (ADFI), an improved overall body weight, andthe ratio of F/G wherein the ratio of F/G is defined as the ADFI/ADG.

The non-human animals may exhibit a decrease in the feed conversionratio of about 2.79% or more as compared to a control group withoutsupplementation of the feed additive composition. The non-human animalsmay exhibit a reduced mortality of at least 10% or more as compared to acontrol group without supplementation of the feed additive composition.The non-human animals may show an improved body weight gain as definedas the percent increase of at least 2.71% as compared to a control groupwithout supplementation of the feed additive composition.

(c) Improved Health

Still another aspect of the disclosure encompasses methods for improvingthe health of non-human animals. “Improved health”, as defined herein,refers to a reduction of incidences of diarrhea, reduction in the numberof days of diarrhea, a decrease in mortality, improving intestinalhealth, reducing microbial pathogens in the intestinal tract of theanimal, a decrease in cytokine panel measuring TNF-alpha, decrease inimmunocrit levels, or combinations thereof in the non-human animals ascompared to a control group.

(d) Reduced Impact of Microbial Pathogens

Still another aspect of the disclosure encompasses methods for reducingthe impact of microbial pathogens in the non-human animals. Reducing theimpact of microbial pathogens may comprise improving the intestinalhealth and the reduction of microbial pathogens in the non-humananimals. Reducing the impact of microbial pathogens may also compriseimproving the general health and performance of non-human animalsexposed to the microbial pathogens.

The term “microbial pathogens”, as defined herein, refers to amicroorganism that has the potential to cause disease. An infection isthe invasion and multiplication of pathogenic microbes in a subject.Disease is when the infection causes damage to the subject's vitalfunctions or systems.

“Improved intestinal health” and the “reduction of microbial pathogens”refer to a reduction in the number of pathogens and a reduction ofinflammation caused by the microbial pathogens in the non-human animalas compared to a control group. Non-limiting examples of pathogenicbacteria that may be controlled using a feed additive composition of thepresent disclosure include Clostridium perfringens, Eimeria maxima,Aeromonas hydrophila, Yersinia enterocolitica, Vibrio spp., Leptospiraspp., Mycobacterium ulcerans, Listeria spp., pathogenic strains of E.coli, Pseudomonas spp. such as aeruginosa, Enterococcus spp., Salmonellaspp., Campylobacter spp., Staphylococcus spp. such as epidermidis, S.aureus (MRSA), M. smegmatis, Streptococcus sp., Clostridia, and M.marinum.

In a preferred alternative of the embodiments, reducing the impact ofmicrobial pathogens comprises reducing the impact of C. perfringens inbroilers. In an alternative of the embodiments, reducing the impact ofmicrobial pathogens comprises controlling necrotic enteritis in thechicken. In another alternative, reducing the impact of microbialpathogens comprises controlling the impact of C. perfringens incombination with a coccidiosis challenge in disease-challenged broilers.

(e) Improved Quality

Another aspect of the disclosure encompasses methods for improving thequality of a product derived from a non-human animal. For instance, whenthe non-human animal is a chicken, an improved breast meat yield,reduced product variability, reduced impact on consumer appeal due topoor color, poor texture, and drip loss in retail display, reducedorganoleptic qualities in cooked chicken including incidence of woodybreast.

Definitions

When introducing elements of the embodiments described herein, thearticles “a”, “an”, “the” and “said” are intended to mean that there areone or more of the elements. The terms “comprising”, “including” and“having” are intended to be inclusive and mean that there may beadditional elements other than the listed elements.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present disclosure shall have the meanings that arecommonly understood by those of ordinary skill in the art. The meaningand scope of the terms should be clear; however, in the event of anylatent ambiguity, definitions provided herein take precedent over anydictionary or extrinsic definition. Further, unless otherwise requiredby context, singular terms as used herein and in the claims shallinclude pluralities, and plural terms shall include the singular.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimits of that range is also specifically disclosed. Each smaller rangebetween any stated value or intervening value in a stated range and anyother stated or intervening value in that stated range is encompassedwithin the invention. The upper and lower limits of these smaller rangescan independently be included or excluded in the range, and each rangewhere either, neither or both limits are included in the smaller rangesis also encompassed within the invention, subject to any specificallyexcluded limit in the stated range. Where the stated range includes oneor both of the limits, ranges excluding either or both of those includedlimits are also included in the invention.

As used herein, the terms “about” and “approximately” designate that avalue is within a statistically meaningful range. Such a range can betypically within 20%, more typically still within 10%, and even moretypically within 5% of a given value or range. The allowable variationencompassed by the terms “about” and “approximately” depends on theparticular system under study and can be readily appreciated by one ofordinary skill in the art.

As used herein, “administering” is used in its broadest sense to meancontacting a non-human animal with a composition disclosed herein.

The phrases “therapeutically effective amount” and “antimicrobialeffective amount” are used interchangeably to mean an amount that isintended to qualify the amount of an agent or compound, that whenadministered, it will achieve the goal of healing an infection site,inhibiting the growth of a microorganism, or otherwise benefiting therecipient environment.

The terms “isolated”, “purified”, or “biologically pure” refer tomaterial that is substantially or essentially free from components thatnormally accompany it as found in its native state. Purity andhomogeneity are typically determined using analytical chemistrytechniques such as polyacrylamide gel electrophoresis or highperformance liquid chromatography. “Purify” or “purification” in otherembodiments means removing at least one contaminant from the compositionto be purified. In this sense, purification does not require that thepurified compound be homogenous, e.g., 100% pure.

As used herein, the terms “treating”, “treatment”, or “to treat” eachmay mean to alleviate, suppress, repress, eliminate, prevent or slow theappearance of symptoms, clinical signs, or underlying pathology of acondition or disorder on a temporary or permanent basis. Preventing acondition or disorder involves administering an agent of the presentinvention to a subject prior to onset of the condition. Suppressing acondition or disorder involves administering an agent of the presentinvention to a subject after induction of the condition or disorder butbefore its clinical appearance. Repressing the condition or disorderinvolves administering an agent of the present invention to a subjectafter clinical appearance of the disease. Prophylactic treatment mayreduce the risk of developing the condition and/or lessen its severityif the condition later develops. For instance, treatment of a microbialinfection may reduce, ameliorate, or altogether eliminate the infection,or prevent it from worsening.

Having described the invention in detail, it will be apparent thatmodifications and variations are possible without departing from thescope of the invention defined in the appended claims.

CITATIONS

-   1. Cobb-Vantress, Inc. 2012. Broiler management guide. Cobb-Vantress    Inc., Siloam Springs, Ark.-   2. Hofacre, C. L., R. Froyman, B. George, M. A. Goodwin, and J.    Brown. 1998. Use of Aviguard, virginiamycin or bacitracin MD against    Clostridium perfringens-associated necrotizing enteritis. J. Appl.    Poult. Res. 7:412-418.

EXAMPLES

The following examples illustrate various embodiments of the invention.

Example 1: Determine the Effect of Feeding Ground Peppers to BroilersChallenged with E. maxima and C. perfringens

This study was conducted to evaluate the effects of ground peppers inbroilers challenged with Eimeria maxima and Clostridium perfringens. Inthis and all subsequent examples, ground peppers refers to encapsulatedground peppers comprising 50% ground peppers and 50% encapsulatingmaterial. A total of 256 day-old male Cobb×Cobb chicks were obtainedfrom a commercial hatchery. Chicks were vaccinated for Marek's andreovirus at the hatchery. The birds were randomly assigned to 32 batterycages with eight chicks per cage. A total of five treatments (Table 1)were allotted, resulting in eight cages per treatment. Temperature andlighting were adjusted according to the husbandry guidelines of the Cobbbroiler (Cobb, 2012). Each cage was equipped with a feed and watertrough. Water and feed were provided ad libitum. The chicks were fed a22.5% crude protein corn soybean-based diet (Table 2) through 28-dpost-hatching. The diet was representative of a local commercialformulation and calculated analyses met or exceeded NRC broiler starterrequirements. Experimental treatment feeds were prepared from thisnon-medicated basal starter feed.

TABLE 1 Experimental treatments Conc. of Treat- Testing Testing E.maxima C. perfringens # of ment product Product Challenge ChallengeCages A No Additive None DOT¹ 14 None 8 B No Additive None DOT 14 DOT19, 20, 8 and 21 C BMD 50.0 g/ton DOT 14 DOT 19, 20, 8 and 21 DEncapsulated  300 g/ton DOT 14 DOT 19, 20, 8 ground and 21 pepper ¹Dayof hatch Encapsulated ground peppers comprise 50% ground peppers and 50%encapsulating material. Capsaicinoid content of encapsulated groundpepper is 0.5%

TABLE 2 Composition of the basal diet¹ Kg/ton Ingredients Corn, YellowGrain 556.29 Soybean Meal Dehulled, Solvent 377.55 Fat, Vegetable 25.55Dicalcium Phosphate 20.39 Calcium Carbonate 10.31 Salt, (NaCl) 4.61Methionine, MHA 3.08 L - LYSINE, 98.5% 0.82 Trace Mineral Premix 0.75Vitamin Premix 0.65 Total 1000.00 Calculated Nutrients Composition CrudeProtein, % 22.45 Fat, % 4.85 Crude Fiber, % 2.19 ME, kcal/Kg 3054.00Lysine, % 1.34 Methionine, % 0.63 Threonine, % 0.92 Calcium, % 0.92Phosphorus, % 0.45 Phosphorus, Available, % 0.67 Ca/P Ratio 2.04 Sodium,% 0.21 ¹Basal diet was prepared by Southern Poultry Research

Bird Allocation and Cage Randomization

Cages were blocked by location in the battery. The study began when thebirds were placed (day of hatch) (DOT 0) at which time they wereallocated to the experimental cages. No birds were replaced during thecourse of the study.

Cage Weights

All birds were weighed on DOT 0, 14, 21, and 28. Feed was weighed on DOT0 and remaining feed was weighed on DOT 14, 21, and 28.

Disease Induction

On DOT 14, all birds were orally inoculated with ˜5,000 oocysts of E.maxima. Starting on DOT 19, all birds, except Treatment 1 (Control) weregiven a broth culture of C. perfringens ˜10⁸ cfu/ml. The birds wereadministered a fresh broth culture once daily for 3 days (on DOTs 19,20, and 21).

Necrotic Enteritis Intestinal Lesion Scoring

On DOT 21, three birds from each cage were selected, sacrificed,weighed, and examined for the degree of presence of Necrotic Enteritislesions. If less than three birds were present at time of scoring, allremaining birds within the cage were scored. The scoring was based on a0 to 3 score, with 0 being normal and 3 being the most severe. (Hofacreet al., 1998).

Management

The facility was checked at least twice daily to assure that all cageshad water and that feed was available in every cage. The birds werewatched closely for any abnormal conditions. The building temperature'srange was maintained at an appropriate temperature for the age of thebirds. Even, continuous light was provided by fluorescent lamps hungvertically along the wall. Feed and water was given ad libitum. Whendead birds were removed from cages, the cage number, date, weight of thebird, sex, and probable cause of death were recorded.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included cage weight gain, feed consumption, feedconversion, necrotic enteritis lesion scores, and % necrotic enteritisrelated mortality were calculated. All results were reported as leastsquares means. The significance level chosen was α=0.05. Treatmenteffect was considered significant if P<0.05, whereas values between0.05≤P≤0.10 were considered as statistical trends.

Results

Broilers fed ground peppers had increased (P<0.05) body weight gainduring the challenge and post challenge periods (d 14-28) compared tothe challenge control birds (Table 3). Birds fed ground peppers hadsimilar body weight gains as the unchallenged controls during thisperiod of age. During all periods, body weight gain was numericallyincreased in birds fed ground peppers compared to the challengecontrols. No significant differences were observed in feed conversionratio (FCR) compared to the challenged control and birds fed groundpeppers. Necrotic enteritis lesion scores and necrotic enteritis relatedmortality were lower (P<0.05) in birds fed the ground peppers comparedto the challenged control birds. No differences were observed betweenthe BMD (bacitracin methylene disalycilate) fed birds and the birds fedground peppers in lesion scores or percent necrotic enteritis relatedmortality, suggesting that the ground peppers was effective incontrolling C. perfringens disease challenge. No differences were notedin feed intake between all treatments.

TABLE 3 Summary 2017 SPRG NQ NE0117 ground peppers Trial Treatment NonChallenged Challenged Challenged Challenged Ground Variable ControlControl BMD peppers PSE P-value No. of Cages 8 8 8 8 No. of Birds 64 6464 64 C. Perfringens Challenge − + + + Live performance, 0-21 d Feedconsumption, Kg 0.77 0.81 0.79 0.81 0.03 0.53 Body weight gain, Kg 0.460.43 0.44 0.43 0.02 0.33 Feed Conversion 1.66^(a) 1.92^(b) 1.79^(ab)1.89^(b) 0.04 0.001 Live performance, 14-21 d Feed consumption, Kg 0.390.42 0.41 0.42 0.02 0.63 Body weight gain, Kg 0.21^(a) 0.14^(b) 0.16^(b)0.16^(b) 0.01 0.0001 Feed Conversion 1.90^(a) 2.99^(c) 2.56^(b)2.67^(bc) 0.12 <.0001 Live performance, 14-28 d Feed consumption, Kg0.88 0.92 0.95 1.07 0.10 0.53 Body weight gain, Kg 0.46^(a) 0.32^(b)0.37^(ab) 0.44^(a) 0.03 0.02 Feed Conversion 1.92^(a) 2.97^(b) 2.58^(b)2.49^(b) 0.15 0.0004 Live performance, 0-28 d Feed consumption, Kg 1.231.18 1.22 1.35 0.07 0.38 Body weight gain, Kg 0.71 0.60 0.65 0.71 0.040.15 Feed Conversion 1.73^(a) 1.96^(b) 1.88^(b) 1.92^(b) 0.04 0.003Necrotic enteritis 0.0^(a) 1.5^(c) 1.1^(b) 1.1^(b) 0.1 <.0001 lesionscore Necrotic enteritis 0.0^(a) 48.4^(c) 28.1^(b) 26.6^(b) 4.0 <.0001related mortality, % ^(a,b,c)Means without a common superscript differ(P < 0.05)

Example 2: Determine the Effect of Feeding Ground Peppers to BroilersChallenged with C. perfringens

This study was conducted to evaluate the effects of ground peppers attwo inclusion levels in broilers raised in pens on new litter and penstop dressed with litter comprising C. perfringens from a previousnecrotic enteritis (NE) challenge study (Table 4). A total of 1200day-old male Ross×Ross chicks were obtained from a commercial hatchery.Chicks were vaccinated for Marek's and reovirus at the hatchery.

TABLE 4 Experimental treatments Litter Starter Grower Finisher # ofTreatment Test product Type DOT¹ 0-13 DOT 14-35 DOT 14-35 Pens AEncapsulated New 0.15 Kg/ton 0.15 Kg/ton 0.15 Kg/ton 8 ground pepper BEncapsulated New 0.3 Kg/ton 0.3 Kg/ton 0.3 Kg/ton 8 ground pepper C NoAdditive New None None None 8 D Encapsulated Reused 0.15 Kg/ton 0.15Kg/ton 0.15 Kg/ton 8 ground pepper E Encapsulated Reused 0.3 Kg/ton 0.3Kg/ton 0.3 Kg/ton 8 ground pepper F No Additive Reused None None None 8¹Day of hatch Capsaicinoid content of encapsulated ground pepper is 0.5%

Experimental Diets

Feed consisted of non-medicated commercial-type broiler starter, grower,and finisher diets compounded according to meet or exceed NRC nutrientguidelines and contained feedstuffs commonly used in the United States(Table 5). Rations were fed ad libitum from date of chick arrival asfollows: Starter—DOT 0 until DOT 15, grower DOT 15 to DOT 35, andfinisher from DOT 35 to DOT 42 (study termination). Diets were fed ascrumbles (starter feed) or pellets (grower and finisher feed). Pelletingof diets (grower and finisher diets) were manufactured with a CaliforniaPellet mill (at 80° C.). Experimental treatment feeds were prepared froma basal feed formulation.

TABLE 5 Composition of the basal diet Starter Grower FinisherIngredients, kg/ton Corn, Yellow Grain 556.29 615.34 640.07 Soybean MealDehulled, Solvent 377.55 318.36 293.02 Fat, Vegetable 25.55 29.11 31.1Dicalcium Phosphate 20.39 18.94 17.86 Calcium Carbonate 10.31 9.85 9.18Salt, (NaCl) 4.61 4.39 4.41 Methionine, MHA 3.08 2.06 2.22 L - LYSINE,98.5% 0.82 0.70 0.89 Trace Mineral Premix 0.75 0.70 0.75 Vitamin Premix0.65 0.50 0.5 Total 1000.00 1000.00 1000.00 Calculated NutrientsComposition Crude Protein, % 22.45 20.0 19.0 Fat, % 4.85 5.35 5.61 CrudeFiber, % 2.19 2.12 2.10 ME, kcal/Kg 3054.00 3125.00 3165.00 Lysine, %1.34 1.16 1.11 Methionine, % 0.63 0.51 0.51 Threonine, % 0.92 0.82 0.77Calcium, % 0.92 0.85 0.80 Phosphorus, % 0.45 0.71 0.68 Phosphorus,Available, % 0.67 0.42 0.40 Ca/P Ratio 2.04 2.02 2.00 Sodium, % 0.210.20 0.20

Animal Information

Upon arrival all birds were vaccinated (by spray cabinet) with acommercially approved coccidia vaccine at the normal recommended dosage.Only healthy chicks were used in this study.

Housing

Chicks were raised in 5×5 feet (1.5 m×1.5 m) floor pens (stockingdensity of 1.0 feet² per bird) in a solid-sided barn, with concretefloors, and under ambient temperature control. At placement, birds wereplaced on reused litter homogenized from a previous necrotic enteritisstudy. Each pen contained 1 (one) tube feeder and six (6) nippledrinkers.

Body and Feed Weight

All birds were weighed by pen on DOT 0, 15, 35, and 42. The trial wasterminated on DOT 42. On DOT 15 remaining starter feed was removed,weighed, and replaced with grower feed to DOT 35. On DOT 35 remaininggrower feed was removed, weighed back, and replaced with finisher feedto DOT 42 (study termination).

Management

All birds were monitored for general flock condition, temperature,lighting, water, feed, litter condition, and unanticipated houseconditions/events. Pens were checked daily for mortality. A grossnecropsy was performed on all dead or culled birds to determine the birdsex and probable cause of death.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included Cage weight gain, feed consumption, feedconversion, percent NE related mortality, and total percent mortality.All results were reported as least squares means. The significance levelchosen was α=0.05. Treatment effect was considered significant ifP<0.05, whereas values between 0.05≤P≤0.10 were considered asstatistical trends.

Results

No performance differences were observed between ground peppersinclusion rates (Table 6). Reused litter decreased (P<0.05) body weightgain and feed intake. Ground peppers tended to reduce feed intake at 15,35, and 42 days of age compared to the no feed additive group. Bodyweight gain tended to increase at 15, 35, and 42 days of age compared tothe control group, and feed conversion was reduced by 0.6 to 2.5% whenground peppers was added to the diets. No mortality differences wereobserved between birds fed ground peppers and the no feed additivegroup.

TABLE 6 Summary 2016 SPRG NQPP102016 Ground peppers 2 levels and 2litter types Treatment Encapsulated Encapsulated EncapsulatedEncapsulated P-value ground pepper ground pepper ground pepper groundpepper Ground Variable 0.15 Kg/ton 0.3 Kg/ton Control 0.15 Kg/ton 0.3Kg/ton Control PSE peppers Litter No. of Pens 8 8 8 8 8 8 No. of Birds200 200 200 200 200 200 Litter Type New New New Reused Reused ReusedLive performance, 15 d Feed consumption, Kg 0.32 0.32 0.32 0.30 0.300.30 0.01 0.90 0.01 Body weight gain, Kg 0.33 0.32 0.32 0.31 0.30 0.290.01 0.24 <.0001 Feed Conversion 1.11 1.14 1.15 1.15 1.16 1.16 0.04 0.540.27 Live performance, 35 d Feed consumption, Kg 2.90 2.88 2.90 2.722.72 2.72 0.03 0.93 <.0001 Body weight gain, Kg 2.07 2.04 2.05 1.99 1.941.97 0.03 0.39 0.0004 Feed Conversion 1.43 (−1.4%) 1.44 (−0.6%) 1.451.39 (−1.4%) 1.43 (−1.4%) 1.41 0.01 0.11 0.003 Live performance, 42 dFeed consumption, Kg 4.01 4.06 4.09 3.91 3.87 3.87 0.05 0.88 <.0001 Bodyweight gain, Kg 2.59 2.61 2.58 2.54 2.50 2.49 0.04 0.69 0.02 FeedConversion 1.58 (−2.5%) 1.58 (−2.5%) 1.62 1.56 (−1.3%) 1.57 (−0.6%) 1.580.02 0.21 0.10 Total Mortality % 3.50 3.00 4.00 2.50 2.50 3.00 1.5 0.630.71

Example 3: Determine the Effect of Feeding Ground Peppers, FormulatedYeast and a Combination of Ground Peppers and Formulated Yeast toBroilers Challenged with C. perfringens

This study was conducted to evaluate the effects of ground peppers andformulated yeast fed to broilers challenged with C. perfringens (Table7). A total of 2000 day-old male Ross×Ross chicks were obtained from acommercial hatchery. Chicks were vaccinated for Marek's and reovirus atthe hatchery.

TABLE 7 Experimental treatments Treat- Starter Grower Finisher # of mentTest product DOT¹ 0-13 DOT 14-35 DOT 14-35 Pens A No Additive None DOT¹14 None 16 B Virginimycin  20 g/ton  20 g/ton  20 g/ton 16 C Formulated0.5 lb/ton 0.5 lb/ton 0.5 lb/ton 16 D yeast 0.3 lb/ton 0.3 lb/ton 0.3lb/ton 16 Encapsulated E ground 0.3 lb/ton + 0.3 lb/ton + 0.3 lb/ton +16 pepper 0.5 lb/ton 0.5 lb/ton 0.5 lb/ton Encapsulated ground pepper +Formulated yeast ¹Day of hatch Capsaicinoid content of encapsulatedground pepper is 0.5%

Experimental Diets

Feed consisted of non-medicated commercial-type broiler starter, grower,and finisher diets compounded according to meet or exceed NRC nutrientguidelines and contained feedstuffs commonly used in the United States(Table 8). Rations were fed ad libitum from date of chick arrival asfollows: Starter—DOT 0 until DOT 15, grower DOT 15 to DOT 35, andfinisher from DOT 35 to DOT 42 (study termination). Diets were fed ascrumbles (starter feed) or pellets (grower and finisher feed). Pelletingof diets (grower and finisher diets) was manufactured with a CaliforniaPellet mill (at 80° C.). Experimental treatment feeds were prepared froma basal feed formulation.

TABLE 8 Composition of the basal diet Starter Grower FinisherIngredients, kg/ton Corn, Yellow Grain 556.29 615.34 640.07 Soybean MealDehulled, Solvent 377.55 318.36 293.02 Fat, Vegetable 25.55 29.11 31.1Dicalcium Phosphate 20.39 18.94 17.86 Calcium Carbonate 10.31 9.85 9.18Salt, (NaCl) 4.61 4.39 4.41 Methionine, MHA 3.08 2.06 2.22 L - LYSINE,98.5% 0.82 0.70 0.89 Trace Mineral Premix 0.75 0.70 0.75 Vitamin Premix0.65 0.50 0.5 Total 1000.00 1000.00 1000.00 Calculated NutrientsComposition Crude Protein, % 22.45 20.0 19.0 Fat, % 4.85 5.35 5.61 CrudeFiber, % 2.19 2.12 2.10 ME, kcal/Kg 3054.00 3125.00 3165.00 Lysine, %1.34 1.16 1.11 Methionine, % 0.63 0.51 0.51 Threonine, % 0.92 0.82 0.77Calcium, % 0.92 0.85 0.80 Phosphorus, % 0.45 0.71 0.68 Phosphorus,Available, % 0.67 0.42 0.40 Ca/P Ratio 2.04 2.02 2.00 Sodium, % 0.210.20 0.20

Animal Information

Upon arrival all birds were vaccinated (by spray cabinet) with acommercially approved coccidia vaccine at the normal recommended dosage.Only healthy chicks were used in this study.

Housing

Chicks were raised in 5×5 feet (1.5 m×1.5 m) floor pens (stockingdensity of 1.0 feet² per bird) in a solid-sided barn, with concretefloors, and under ambient temperature control. At placement, birds wereplaced on reused litter homogenized from a previous necrotic enteritisstudy. Each pen contained 1 (one) tube feeder and six (6) nippledrinkers.

C. perfringens Application

Feed in each pen had fifty (50) ml applied at 15 days of ˜1×10⁸ CFU/mlper pen of C.P#6.

Body and Feed Weight

All birds were weighed by pen on DOT 0, 15, 35, and 42. The trial wasterminated on DOT 42. On DOT 15 remaining starter feed was removed,weighed, and replaced with grower feed to DOT 35. On DOT 35 remaininggrower feed was removed, weighed back, and replaced with finisher feedto DOT 42 (study termination).

Management

All birds were monitored for general flock condition, temperature,lighting, water, feed, litter condition, and unanticipated houseconditions/events. Pens were checked daily for mortality. A grossnecropsy was performed on all dead or culled birds to determine the birdsex and probable cause of death.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Pen served as the experimental unit.The statistical model included pen weight gain, feed consumption, feedconversion, percent NE related mortality, and total percent mortality.All results were reported as least squares means. The significance levelchosen was α=0.05. Treatment effect was considered significant ifP<0.05, whereas values between 0.05≤P≤0.10 were considered asstatistical trends.

Results

Broilers fed the combination of ground peppers and formulated yeast hada decreased (P<0.05) feed intake compared to birds fed the groundpeppers (Table 9). Feed conversion tended (P=0.08) to decrease in birdsfed the combination of ground peppers and formulated yeast compared tothe ground peppers fed birds at 15 d of age. Body weight and FCR wasnumerically improved in birds fed the combination of ground peppers andformulated yeast compared to formulated yeast and ground peppers fedtreatments. Necrotic enteritis related mortality was numerically lowerfor the combination feed treatment compared to the control andvirginiamycin fed birds.

TABLE 9 Summary 2017 Ground peppers and Formulated Yeast Trial TreatmentGround peppers + Formulated Ground Formulated Variable ControlVirginiamycin yeast peppers yeast PSE P-value No. of Pens 16 No. ofBirds 400 Live performance, 15 d Feed consumption, Kg 0.44^(ab)0.44^(ab) 0.43^(ab) 0.45^(a) 0.41^(b) 0.01 0.04 Body weight gain, Kg0.37 0.38 0.37 0.37 0.37 0.01 0.63 Feed Conversion 1.35^(xy) 1.32^(xy)1.32^(xy) 1.38^(xy) 1.23^(y) 0.04 0.08 Live performance, 35 d Feedconsumption, Kg 2.63 2.62 2.59 2.68 2.59 0.06 0.83 Body weight gain, Kg1.61^(x) 1.57^(xy) 1.54^(y) 1.57^(xy) 1.60^(xy) 0.02 0.08 FeedConversion 1.69 1.73 1.73 1.77 1.67 0.05 0.31 Live performance, 42 dFeed consumption, Kg 3.87 3.70 3.85 3.87 3.81 0.09 0.63 Body weightgain, Kg 2.24 2.17 2.18 2.18 2.21 0.02 0.12 Feed Conversion 1.77 1.761.81 1.82 1.77 0.04 0.56 Necrotic enteritis 3.50 3.50 3.75 3.00 4.251.15 0.49 related mortality, % Total Mortality % 0.63 0.50 0.38 1.250.38 0.32 0.28 ^(a,b,c)Means without a common superscript differ (P <0.05) ^(x,y)Means without a common superscript tend to differ (P < 0.10)

Example 4: Determine the Effect of Feeding Ground Peppers at FourInclusion Levels Fed in Combination with Formulated Yeast Challengedwith C. perfringens

This study was conducted to evaluate the effects of ground peppers fedat four inclusion levels and formulated yeast fed to broilers challengedwith C. perfringens (Table 10). A total of 3000 day-old male Ross×Rosschicks were obtained from a commercial hatchery. Chicks were vaccinatedfor Marek's and reovirus at the hatchery.

TABLE 10 Experimental treatments Treat- Starter Grower Finisher # ofment Test product DOT¹ 0-15 DOT 15-28 DOT 28-42 Pens A No Additive NoneDOT¹ 14 None 20 B Virginimycin 20 g/ton 20 g/ton 20 g/ton 20 CEncapsulated 0.1 lb/ton + 0.1 lb/ton + 0.1 lb/ton + 20 ground 0.50lb/ton 0.50 lb/ton 0.50 lb/ton pepper + Formulated yeast D Encapsulated0.3 lb/ton + 0.3 lb/ton + 0.3 lb/ton + 20 ground 0.50 lb/ton 0.50 lb/ton0.50 lb/ton pepper + Formulated yeast E Encapsulated 0.5 lb/ton + 0.5lb/ton + 0.5 lb/ton + 20 ground 0.50 lb/ton 0.50 lb/ton 0.50 lb/tonpepper + Formulated yeast F Encapsulated 0.70 lb/ton + 0.70 lb/ton +0.70 lb/ton + 20 ground 0.50 lb/ton 0.50 lb/ton 0.50 lb/ton pepper +Formulated yeast ¹Day of hatch Capsaicinoid content of encapsulatedground pepper is 0.5%

Experimental Diets

Feed consisted of non-medicated commercial-type broiler starter, grower,and finisher diets compounded according to meet or exceed NRC nutrientguidelines and contained feedstuffs commonly used in the United States(Table 11). Rations were fed ad libitum from date of chick arrival asfollows: Starter—DOT 0 until DOT 15, grower DOT 15 to DOT 28, andfinisher from DOT 28 to DOT 42 (study termination). Diets were fed ascrumbles (starter feed) or pellets (grower and finisher feed). Pelletingof diets (grower and finisher diets) was manufactured with a CaliforniaPellet mill (at 80° C.). Experimental treatment feeds were prepared froma basal feed formulation.

TABLE 11 Composition of the basal diet Ingredients, kg/ton StarterGrower Finisher Corn, Yellow Grain 556.29 615.34 640.07 Soybean MealDehulled, Solvent 377.55 318.36 293.02 Fat, Vegetable 25.55 29.11 31.1Dicalcium Phosphate 20.39 18.94 17.86 Calcium Carbonate 10.31 9.85 9.18Salt, (NaCl) 4.61 4.39 4.41 Methionine, MHA 3.08 2.06 2.22 L—LYSINE,98.5% 0.82 0.70 0.89 Trace Mineral Premix 0.75 0.70 0.75 Vitamin Premix0.65 0.50 0.5 Total 1000.00 1000.00 1000.00 Calculated NutrientsComposition Crude Protein, % 22.45 20.0 19.0 Fat, % 4.85 5.35 5.61 CrudeFiber, % 2.19 2.12 2.10 ME, kcal/Kg 3054.00 3125.00 3165.00 Lysine, %1.34 1.16 1.11 Methionine, % 0.63 0.51 0.51 Threonine, % 0.92 0.82 0.77Calcium, % 0.92 0.85 0.80 Phosphorus, % 0.45 0.71 0.68 Phosphorus,Available, % 0.67 0.42 0.40 Ca/P Ratio 2.04 2.02 2.00 Sodium, % 0.210.20 0.20

Animal Information

Upon arrival all birds were vaccinated (by spray cabinet) with acommercially approved coccidia vaccine at the normal recommended dosage.Only healthy chicks were used in this study.

Housing

Chicks were raised in 5×5 feet (1.5 m×1.5 m) floor pens (stockingdensity of 1.0 feet² per bird) in a solid-sided barn, with concretefloors, and under ambient temperature control. At placement, birds wereplaced on reused litter homogenized from a previous necrotic enteritisstudy. Each pen contained 1 (one) tube feeder and six (6) nippledrinkers.

C. perfringens Application

Feed in each pen had fifty (50) ml applied at 15 days of ˜1×10⁸ CFU/mlper pen of C.P#6.

Body and Feed Weight

All birds were weighed by pen on DOT 0, 15, 35, and 42. The trial wasterminated on DOT 42. On DOT 15 remaining starter feed was removed,weighed, and replaced with grower feed to DOT 35. On DOT 35 remaininggrower feed was removed, weighed back, and replaced with finisher feedto DOT 42 (study termination).

Management

All birds were weighed by pen on DOT 0, 15, 28, and 42. The trial wasterminated on DOT 42. On DOT 15 remaining starter feed was removed,weighed, and replaced with grower feed to DOT 28. On DOT 28 remaininggrower feed was removed, weighed back, and replaced with finisher feedto DOT 42 (study termination).

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Pen served as the experimental unit.The statistical model included pen weight gain, feed consumption, feedconversion, percent NE related mortality, and total percent mortality.All results were reported as least squares means. The significance levelchosen was α=0.05. Treatment effect was considered significant ifP<0.05, whereas values between 0.05≤P≤0.10 were considered asstatistical trends.

Results

The combination of ground peppers and formulated yeast improved bodyweight gain, FCR and survivability (Table 12). Broilers fed thecombination of ground peppers at 0.3 lb/ton finished feed and formulatedyeast had a decreased (P<0.05) FCR at 15, 28 and 42 days of age comparedto birds fed no feed additive. Body weight gain was increased (P<0.05)in birds fed the 0.3 lb ground peppers/ton plus formulated yeastcompared to the control fed birds at 28 days of age. Birds fed theground peppers at 0.7 lb/ton finished feed tended to have higher(P=0.07) feed intake compared to the 0.3 lb ground peppers/ton group.Total mortality was numerically reduced in birds fed all feed additivescompared to the no feed additive fed group. These results suggest thatfeeding broilers ground peppers especially at the 0.3 lb/ton inclusionrate with formulated yeast is effective in controlling C. perfringensdisease challenge.

TABLE 12 Summary 2017 SPRG NQPYC052017-48 Ground peppers and FormulatedYeast Trial Treatment Encapsulated Encapsulated EncapsulatedEncapsulated ground pepper + ground pepper + ground pepper + groundpepper + Formulated Formulated Formulated Formulated Variable ControlVirginiamycin yeast yeast yeast yeast PSE P-value No. of Pens 20 20 2020 20 20 No. of Birds 500 500 500 500 500 500 Inclusion Rate None 20g/ton 0.10 + 0.50 0.30 + 0.50 0.50 + 0.50 0.70 + 0.50 lb/ton lb/tonlb/ton lb/ton Live performance, 15 d Feed consumption, Kg 0.40 0.39 0.390.39 0.40 0.41 0.01 0.16 Body weight gain, Kg 0.32 0.33 0.32 0.33 0.330.33 0.01 0.23 Feed Conversion 1.40^(a) 1.33^(b) 1.35^(ab) 1.32^(b)1.35^(ab) 1.36^(ab) 0.03 0.02 Live performance, 35 d Feed consumption,Kg 1.76^(xy) 1.73^(x) 1.73^(x) 1.74^(xy) 1.73^(x) 1.81^(y) 0.03 0.06Body weight gain, Kg 1.08^(a) 1.10^(ab) 1.07^(ab) 1.12^(b) 1.11^(ab)1.10^(ab) 0.02 0.04 Feed Conversion 1.69^(a) 1.62^(abc) 1.66^(abc)1.59^(c) 1.61^(bc) 1.69^(a) 0.04 0.003 Live performance, 42 d Feedconsumption, Kg 3.90^(xy) 3.90^(xy) 3.89^(xy) 3.84^(x) 3.90^(xy)3.98^(y) 0.04 0.07 Body weight gain, Kg 2.21 2.27 2.22 2.27 2.29 2.270.03 0.12 Feed Conversion 1.79^(a) 1.74^(abc) 1.78^(b) 1.72^(c)1.73^(bc) 1.77^(abc) 0.02 0.005 Total Mortality % 6.29 5.36 5.40 3.604.80 5.00 1.49 0.61 ^(a,b,c)Means without a common superscript differ (P< 0.05) ^(x,y)Means without a common superscript tend to differ (P <0.10)

Example 5: Determine the Effect of Feeding Capsicum Product, FormulatedYeast and Bacillus licheniformis to Broilers Challenged with E. maximaand C. perfringens

A total of 256-day-old male Cobb×Cobb chicks were obtained from acommercial hatchery. Chicks were vaccinated for Marek's disease andreovirus at the hatchery. The birds were randomly assigned to 32 batterycages with eight chicks per cage. A total of four treatments (Table 13)were allotted resulting in eight pens per treatment. Temperature andlighting were adjusted according to the husbandry guidelines of the Cobbbroiler (Cobb, 2012). Each cage was equipped with a feed and watertrough. Water and feed were provided ad libitum. A 22.5% crude proteincorn soybean-based diet (Table 14) was fed through 28-d posthatching.The diet was representative of a local commercial formulation andcalculated analyses met or exceeded NRC broiler starter requirements.Experimental treatment feeds were prepared from this non-medicated basalstarter feed.

Bird Allocation and Cage Randomization

Cages were blocked by location in the battery. The study began when thebirds were placed (day of hatch) (DOT 0) at which time they wereallocated to the experimental cages. No birds were replaced during thecourse of the study.

Cage Weights

All birds were weighed on DOT 0, 14, 21, and 28. Feed was weighed on DOT0 and remaining feed was weighed on DOT 14, 21, and 28.

Disease Induction

On DOT 14, all birds were orally inoculated with ˜5,000 oocysts of E.maxima. Starting on DOT 19, all birds, except Treatment 1 (Control) weregiven a broth culture of C. perfringens ˜108 cfu/ml. The birds wereadministered a fresh broth culture once daily for 3 days (on DOTs 19,20, and 21).

Necrotic Enteritis Intestinal Lesion Scoring

On DOT 21, three birds from each cage were selected, sacrificed,weighed, and examined for the degree of presence of Necrotic Enteritislesions. If less than three birds were present at time of scoring, allremaining birds within the cage were scored. The scoring was based on a0 to 3 score, with 0 being normal and 3 being the most severe. (Hofacreet. al, 1998)

Management

The facility was checked at least twice daily to assure that all cageshad water and that feed was available in every cage. The buildingtemperature's range was maintained at an appropriate temperature for theage of the birds, and continuous light was provided by fluorescent lampshung vertically along the wall. Feed and water were given ad libitum.When mortality birds were removed from cages, the cage number, date,weight of the bird, sex, and probable cause of death were recorded.

Data Analysis

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included cage weight gain, feed consumption, feedconversion, necrotic enteritis lesion scores, and % necrotic enteritisrelated mortality were calculated. All results were reported as leastsquares means. The significance level chosen was α=0.05. Treatmenteffect was considered significant if P<0.05, whereas values between0.05≤P≤0.10 were considered as statistical trends.

Summary

Broilers fed the combination of Capsicum Product, formulated yeast andB. licheniformis had similar (P>0.10) FCR to 21 days of age as theunchallenged no feed additive treatment group. Broilers fed thecombination of Capsicum Product, formulated yeast and B. licheniformisand BMD had decreased (P<0.05) FCR during the challenge and postchallenge periods (d 14-28) compared to the challenge control birds(Table 15). Overall FCR was reduced (P<0.05) in birds fed BMD and thecombination products compared to the challenged no feed additivecontrol. Birds fed the combination of Capsicum Product, formulated yeastand B. licheniformis had similar (P>0.10) d 1-28 FCR to the unchallengedcontrol group. Body weight gain was not impacted by either the BMD orcombination treatment compared to the challenged no feed additivecontrol group. No differences were noted in feed intake between alltreatments. Necrotic enteritis lesion scores and necrotic enteritisrelated mortality were lower (P<0.05) in birds fed BMD and CapsicumProduct, formulated yeast and B. licheniformis treatment combinationcompared to the challenged control birds. The data indicates that thetreatment combination of Capsicum Product, formulated yeast and B.licheniformis and BMD are effective in controlling C. perfringensdisease challenge.

TABLE 13 Experimental treatments Conc. of Testing E. maxima C.perfringens # of Treatment Testing Product Product Challenge ChallengeCages A No Additive None DOT¹ 14 None 8 B No Additive None DOT 14 DOT19, 20, and 21 8 C BMD 50.0 g/ton DOT 14 DOT 19, 20, and 21 8 D Capsicum0.30 lb/ton + 0.5 lb/ DOT 14 DOT 19, 20, and 21 8 Product + ton + 3 ×10⁵ cfu/g formulated yeast + in final feed B. licheniformis ¹Day ofhatch

TABLE 1.14 Composition of the basal diet¹ Ingredients Kg/ton Corn,Yellow Grain 556.29 Soybean Meal Dehulled, 377.55 Solvent Fat, Vegetable25.55 Dicalcium Phosphate 20.39 Calcium Carbonate 10.31 Salt, (NaCl)4.61 Methionine, MHA 3.08 L—LYSINE, 98.5% 0.82 Trace Mineral Premix 0.75Vitamin Premix 0.65 Total 1000.00 Calculated Nutrients Composition CrudeProtein, % 22.45 Fat, % 4.85 Crude Fiber, % 2.19 ME, kcal/Kg 3054.00Lysine, % 1.34 Methionine, % 0.63 Threonine, % 0.92 Calcium, % 0.92Phosphorus, % 0.45 Phosphorus, Available, % 0.67 Ca/P Ratio 2.04 Sodium,% 0.21 ¹Basal diet was prepared by Southern Poultry Research

TABLE 15 Summary 2018 SPRG NQ NE0218 Trial Treatment Non CapsicumProduct + Challenged Challenged formulated yeast + Variable ControlControl BMD B. licheniformis No. of Cages 8   8   8   8   No. of Birds64    64    64    64    C. Perfringens − + + + Challenge Liveperformance, 0-21 d Feed consumption, Kg 0.96 0.95 0.94 0.92 Body weightgain, Kg  0.57^(a)  0.51^(b)  0.52^(ab)  0.52^(ab) Feed Conversion 1.69^(b)  1.85^(a)  1.81^(a)  1.76^(ab) Live performance, 14-21 d Feedconsumption, Kg 0.50 0.51 0.49 0.48 Body weight gain, Kg  0.31^(a) 0.26^(b)  0.28^(b)  0.26^(b) Feed Conversion  1.62^(c)  1.98^(a) 1.77^(b)  1.81^(b) Live performance, 14-28 d Feed consumption, Kg 0.901.08 0.91 0.92 Body weight gain, Kg 0.58 0.53 0.53 0.51 Feed Conversion1.56^(c)  2.04^(a)  1.73^(bc)  1.79^(b) Live performance, 0-28 d Feedconsumption, Kg 1.37 1.50 1.37 1.35 Body weight gain, Kg 0.84 0.79 0.770.77 Feed Conversion 1.63^(c)  1.90^(a)  1.78^(b)  1.75^(bc) Necroticenteritis lesion 0.0^(c ) 0.9^(a ) 0.5^(b ) 0.4^(b ) score Necroticenteritis related 0^(b )  11^(a )  2^(b )  3^(b )  mortality, %^(a,b,c)Means within a row with no common superscripts differsignificantly (P < 0.05)

Example 6: Determine the Effect of Feeding Antimicrobial Clay andBacillus licheniformis to Broilers Challenged with E. maxima and C.perfringens

A total of 256-day-old male Cobb×Cobb chicks were obtained from acommercial hatchery. Chicks were vaccinated for Marek's disease andreovirus at the hatchery. The birds were randomly assigned to 32 batterycages with eight chicks per cage. A total of four treatments (Table 16)were allotted resulting in eight pens per treatment. Temperature andlighting were adjusted according to the husbandry guidelines of the Cobbbroiler (Cobb, 2012). Each cage was equipped with a feed and watertrough. Water and feed were provided ad libitum. A 22.5% crude proteincorn soybean-based diet (Table 17) through 28-d posthatching. The dietwas representative of a local commercial formulation and calculatedanalyses met or exceeded NRC broiler starter requirements. Experimentaltreatment feeds were prepared from this non-medicated basal starterfeed.

Bird Allocation and Cage Randomization

Cages were blocked by location in the battery. The study began when thebirds were placed (day of hatch) (DOT 0) at which time they wereallocated to the experimental cages. No birds were replaced during thecourse of the study.

Cage Weights

All birds were weighed on DOT 0, 14, 21, and 28. Feed was weighed on DOT0 and remaining feed was weighed on DOT 14, 21, and 28.

Disease Induction

On DOT 14, all birds were orally inoculated with ˜5,000 oocysts of E.maxima.

Starting on DOT 19, all birds, except Treatment 1 (Control) were given abroth culture of C. perfringens ˜10⁸ cfu/ml. The birds were administereda fresh broth culture once daily for 3 days (on DOTs 19, 20, and 21).

Necrotic Enteritis Intestinal Lesion Scoring

On DOT 21, three birds from each cage were selected, sacrificed,weighed, and examined for the degree of presence of Necrotic Enteritislesions. If less than three birds were present at time of scoring, allremaining birds within the cage were scored. The scoring was based on a0 to 3 score, with 0 being normal and 3 being the most severe. (Hofacreet. al, 1998)

Management

The facility was checked at least twice daily to assure that all cageshad water and that feed was available in every cage. The birds werewatched closely for any abnormal conditions. The building temperature'srange was maintained at an appropriate temperature for the age of thebirds. Even, continuous light was provided by fluorescent lamps hungvertically along the wall. Feed and water were given ad libitum. Whenmortality birds were removed from cages, the cage number, date, weightof the bird, sex, and probable cause of death were recorded.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included cage weight gain, feed consumption, feedconversion, necrotic enteritis lesion scores, and % necrotic enteritisrelated mortality were calculated. All results were reported as leastsquares means. The significance level chosen was α=0.05. Treatmenteffect was considered significant if P<0.05, whereas values between0.05≤P≤0.10 were considered as statistical trends.

Summary

Broilers fed the combination of Antimicrobial Clay and B. licheniformisand BMD had increased (P<0.05) body weight gain and decreased (P<0.05)FCR during the challenge and post challenge periods (d 14-28) comparedto the challenge control birds (Table 18). Similarly, overall (d 0-28)FCR was reduced and body weight gain was increased in birds fed theAntimicrobial Clay and B. licheniformis combination and BMD compared tothe challenged control birds and similar (P>0.10) FCR and body weightgain to the unchallenged control group.

Necrotic enteritis lesion score was lower (P<0.05) in birds fed theAntimicrobial Clay and B. licheniformis combination treatment and BMDcompared to the challenged control birds. Birds fed the BMD feedtreatment had lower (P<0.05) necrotic enteritis mortality compared tothe challenged control group. Birds fed Antimicrobial Clay and B.licheniformis had similar (P>0.10) mortality compared to the BMD andchallenged control groups. The data suggests that both the combinationAntimicrobial Clay and B. licheniformis product and BMD were effectivein controlling C. perfringens disease challenge.

TABLE 16 Experimental treatments Conc. of Testing E. maxima C.perfringens # of Treatment Testing Product Product Challenge ChallengeCages A No Additive None DOT¹ 14 None 8 B No Additive None DOT 14 DOT19, 20, and 21 8 C BMD 50.0 g/ton DOT 14 DOT 19, 20, and 21 8 DAntimicrobial 4.0 lb/ton + 3 × 10⁵ cfu/g DOT 14 DOT 19, 20, and 21 8Clay + B. licheniformis in final feed ¹Day of hatch

TABLE 17 Composition of the basal diet¹ Ingredients Kg/ton Corn, YellowGrain 556.29 Soybean Meal Dehulled, 377.55 Solvent Fat, Vegetable 25.55Dicalcium Phosphate 20.39 Calcium Carbonate 10.31 Salt, (NaCl) 4.61Methionine, MHA 3.08 L—LYSINE, 98.5% 0.82 Trace Mineral Premix 0.75Vitamin Premix 0.65 Total 1000.00 Calculated Nutrients Composition CrudeProtein, % 22.45 Fat, % 4.85 Crude Fiber, % 2.19 ME, kcal/Kg 3054.00Lysine, % 1.34 Methionine, % 0.63 Threonine, % 0.92 Calcium, % 0.92Phosphorus, % 0.45 Phosphorus, Available, % 0.67 Ca/P Ratio 2.04 Sodium,% 0.21 ¹Basal diet was prepared by Southern Poultry Research

TABLE 18 Summary 2018 SPRG NQ NE0718 Trial Treatment Non ChallengedChallenged Antimicrobial Clay + Variable Control Control BMD B.licheniformis No. of Cages 8 8 8 8 No. of Birds 64 64 64 64 C.Perfringens − + + + Challenge Live performance, 0-21 d Feed 0.86 0.930.87 0.88 consumption, Kg Body weight 0.56^(a) 0.51^(b) 0.57^(a)0.55^(ab) gain, Kg Feed 1.53^(c) 1.82^(a) 1.53^(c) 1.61^(bc) ConversionLive performance, 14-21 d Feed 0.52 0.51 0.55 0.55 consumption, Kg Bodyweight 0.34^(a) 0.24^(c) 0.32^(ab) 0.30^(b) gain, Kg Feed 1.53^(c)2.12^(a) 1.72^(b) 1.82^(b) Conversion Live performance, 14-28 d Feed0.87^(a) 0.79^(b) 0.98^(a) 0.92^(a) consumption, Kg Body weight 0.56^(a)0.38^(b) 0.56^(a) 0.53^(a) gain, Kg Feed 1.56^(c) 2.09^(a) 1.75^(b)1.73^(b) Conversion Live performance, 0-28 d Feed 1.21 1.19 1.30 1.26consumption, Kg Body weight 0.78^(a) 0.64^(b) 0.81^(a) 0.79^(a) gain, KgFeed 1.55^(b) 1.86^(a) 1.60^(b) 1.60^(b) Conversion Necrotic enteritis0.00^(c) 1.17^(a) 0.79^(b) 0.58^(b) lesion score Necrotic enteritis0.0^(b) 4.7^(a) 0.0^(b) 1.56^(ab) related mortality, % ^(a,b,c)Meanswithin a row with no common superscripts differ significantly (P < 0.05)

Example 7: Determine the Effect of Feeding Capsicum Product and Bacilluslicheniformis to Broilers Challenged with E. maxima and C. perfringens

A total of 256-day-old male Cobb×Cobb chicks were obtained from acommercial hatchery. Chicks were vaccinated for Marek's disease andreovirus at the hatchery. The birds were randomly assigned to 32 batterycages with eight chicks per cage. A total of four treatments (Table 19)were allotted resulting in eight pens per treatment. Temperature andlighting were adjusted according to the husbandry guidelines of the Cobbbroiler (Cobb, 2012). Each cage was equipped with a feed and watertrough. Water and feed were provided ad libitum. A 22.5% crude proteincorn soybean-based diet (Table 20) through 28-d posthatching. The dietwas representative of a local commercial formulation and calculatedanalyses met or exceeded NRC broiler starter requirements. Experimentaltreatment feeds were prepared from this non-medicated basal starterfeed.

Bird Allocation and Cage Randomization

Cages were blocked by location in the battery. The study began when thebirds were placed (day of hatch) (DOT 0) at which time they wereallocated to the experimental cages. No birds were replaced during thecourse of the study.

Cage Weights

All birds were weighed on DOT 0, 14, 21, and 28. Feed was weighed on DOT0 and remaining feed was weighed on DOT 14, 21, and 28.

Disease Induction

On DOT 14, all birds were orally inoculated with ˜5,000 oocysts of E.maxima.

Starting on DOT 19, all birds, except Treatment 1 (Control) were given abroth culture of C. perfringens ˜10⁸ cfu/ml. The birds were administereda fresh broth culture once daily for 3 days (on DOTs 19, 20, and 21).

Necrotic Enteritis Intestinal Lesion Scoring

On DOT 21, three birds from each cage were selected, sacrificed,weighed, and examined for the degree of presence of Necrotic Enteritislesions. If less than three birds were present at time of scoring, allremaining birds within the cage were scored. The scoring was based on a0 to 3 score, with 0 being normal and 3 being the most severe. (Hofacreet. al, 1998)

Management

The facility was checked at least twice daily to assure that all cageshad water and that feed was available in every cage. The birds werewatched closely for any abnormal conditions. The building temperature'srange was maintained at an appropriate temperature for the age of thebirds. Even, continuous light was provided by fluorescent lamps hungvertically along the wall. Feed and water were given ad libitum. Whenmortality birds were removed from cages, the cage number, date, weightof the bird, sex, and probable cause of death were recorded.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included cage weight gain, feed consumption, feedconversion, necrotic enteritis lesion scores, and % necrotic enteritisrelated mortality were calculated. All results were reported as leastsquares means. The significance level chosen was α=0.05. Treatmenteffect was considered significant if P<0.05, whereas values between0.05≤P≤0.10 were considered as statistical trends.

Summary

Broilers fed the combination of Capsicum Product and B. licheniformisand BMD had increased (P<0.05) body weight gain and decreased (P<0.05)FCR during the challenge and post challenge periods (d 14-28) comparedto the challenge control birds (Table 21). Similarly, overall (d 0-28)body weight gain was increased in birds fed BMD compared to thechallenged control birds and similar (P>0.10) to the unchallengedcontrol group. Broilers fed the combination of Capsicum Product and B.licheniformis had increased (P<0.05) body weight gain compared to thechallenged no feed additive group.

Necrotic enteritis lesion score was lower (P<0.05) in birds fed theCapsicum Product and B. licheniformis combination treatment and BMDcompared to the challenged control birds. No differences (P<0.10)necrotic enteritis mortality were noted. The data suggests that both thecombination Capsicum Product and B. licheniformis product and BMD wereeffective in controlling C. perfringens disease challenge.

TABLE 19 Experimental treatments Testing Conc. of Testing E. maxima C.perfringens # of Treatment Product Product Challenge Challenge Cages ANo Additive None DOT¹ 14 None 8 B No Additive None DOT 14 DOT 19, 20,and 21 8 C BMD 50.0 g/ton DOT 14 DOT 19, 20, and 21 8 D Capsicum 0.3lb/ton + 3 × 10⁵ cfu/g DOT 14 DOT 19, 20, and 21 8 Product + in finalfeed B. licheniformis ¹Day of hatch

TABLE 20 Composition of the basal diet¹ Ingredients Kg/ton Corn, YellowGrain 556.29 Soybean Meal Dehulled, 377.55 Solvent Fat, Vegetable 25.55Dicalcium Phosphate 20.39 Calcium Carbonate 10.31 Salt, (NaCl) 4.61Methionine, MHA 3.08 L—LYSINE, 98.5% 0.82 Trace Mineral Premix 0.75Vitamin Premix 0.65 Total 1000.00 Calculated Nutrients Composition CrudeProtein, % 22.45 Fat, % 4.85 Crude Fiber, % 2.19 ME, kcal/Kg 3054.00Lysine, % 1.34 Methionine, % 0.63 Threonine, % 0.92 Calcium, % 0.92Phosphorus, % 0.45 Phosphorus, Available, % 0.67 Ca/P Ratio 2.04 Sodium,% 0.21 ¹Basal diet was prepared by Southern Poultry Research

TABLE 21 Summary 2018 SPRG NO NE0818 Trial Treatment Non ChallengedChallenged Capsicum Product + Variable Control Control BMD B.licheniformis No. of Cages 8 8 8 8 No. of Birds 64 64 64 64 C.Perfringens − + + + Challenge Live performance, 0-21 d Feed 1.04 0.941.07 1.01 consumption, Kg Body weight 0.72^(a) 0.54^(b) 0.66^(a)0.60^(b) gain, Kg Feed 1.45^(b) 1.74^(a) 1.62^(ab) 1.69^(a) ConversionLive performance, 14-21 d Feed 0.59 0.52 0.58 0.54 consumption, Kg Bodyweight 0.39^(a) 0.21^(c) 0.30^(b) 0.27^(b) gain, Kg Feed 1.52^(c)2.47^(a) 1.95^(b) 2.01^(b) Conversion Live performance, 14-28 d Feed0.99 0.87 1.08 0.97 consumption, Kg Body weight 0.63^(a) 0.39^(c)0.56^(ab) 0.50^(b) gain, Kg Feed 1.57^(c) 2.23^(a) 1.93^(b) 1.94^(b)Conversion Live performance, 0-28 d Feed 1.43 1.27 1.54 1.42consumption, Kg Body weight 0.96^(a) 0.72^(c) 0.92^(ab) 0.83^(b) gain,Kg Feed 1.49^(b) 1.77^(a) 1.67^(a) 1.71^(a) Conversion Necroticenteritis 0.00^(c) 1.04^(a) 0.50^(b) 0.42^(b) lesion score Necroticenteritis 0.0 1.6 0.0 0.0 related mortality, % ^(a,b,c)Means within arow with no common superscripts differ significantly (P < 0.05)

Example 8: Determine the Effect of Feeding Formulated Yeast and Bacilluslicheniformis to Broilers Challenged with E. maxima and C. perfringens

A total of 256-day-old male Cobb×Cobb chicks were obtained from acommercial hatchery. Chicks were vaccinated for Marek's and reovirus atthe hatchery. The birds were randomly assigned to 32 battery cages witheight chicks per cage. A total of four treatments (Table 22) wereallotted resulting in eight pens per treatment. Temperature and lightingwere adjusted according to the husbandry guidelines of the Cobb broiler(Cobb, 2012). Each cage was equipped with a feed and water trough. Waterand feed were provided ad libitum. A 22.5% crude protein cornsoybean-based diet (Table 23) through 28-d posthatching. The diet wasrepresentative of a local commercial formulation and calculated analysesmet or exceeded NRC broiler starter requirements. Experimental treatmentfeeds were prepared from this non-medicated basal starter feed.

Bird Allocation and Cage Randomization

Cages were blocked by location in the battery. The study began when thebirds were placed (day of hatch) (DOT 0) at which time they wereallocated to the experimental cages. No birds were replaced during thecourse of the study.

Cage Weights

All birds were weighed on DOT 0, 14, 21, and 28. Feed was weighed on DOT0 and remaining feed was weighed on DOT 14, 21, and 28.

Disease Induction

On DOT 14, all birds were orally inoculated with ˜5,000 oocysts of E.maxima.

Starting on DOT 19, all birds, except Treatment 1 (Control) were given abroth culture of C. perfringens ˜10⁸ cfu/ml. The birds were administereda fresh broth culture once daily for 3 days (on DOTs 19, 20, and 21).

Necrotic Enteritis Intestinal Lesion Scoring

On DOT 21, three birds from each cage were selected, sacrificed,weighed, and examined for the degree of presence of Necrotic Enteritislesions. If less than three birds were present at time of scoring, allremaining birds within the cage were scored. The scoring was based on a0 to 3 score, with 0 being normal and 3 being the most severe. (Hofacreet. al, 1998)

Management

The facility was checked at least twice daily to assure that all cageshad water and that feed was available in every cage. The birds werewatched closely for any abnormal conditions. The building temperature'srange was maintained at an appropriate temperature for the age of thebirds. Even, continuous light was provided by fluorescent lamps hungvertically along the wall. Feed and water were given ad libitum. Whenmortality birds were removed from cages, the cage number, date, weightof the bird, sex, and probable cause of death were recorded.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Cage served as the experimental unit.The statistical model included cage weight gain, feed consumption, feedconversion, necrotic enteritis lesion scores, and % necrotic enteritisrelated mortality were calculated. All results were reported as leastsquares means. The significance level chosen was α=0.05. Treatmenteffect was considered significant if P<0.05, whereas values between0.05≤P≤0.10 were considered as statistical trends.

Summary

Broilers fed the combination of formulated yeast and B. licheniformishad increased (P<0.05) body weight gain and decreased (P<0.05) FCRduring the challenge and post challenge periods (d 14-28) compared tothe challenge control birds (Table 24). During this period birds fed BMDhad similar body weight gains and decreased (P<0.05) FCR compared to thechallenged control. Overall (d 0-28) FCR was decreased (P<0.05) in birdsfed the combination of formulated yeast and B. licheniformis and BMDcompared to the challenged control birds and similar to the unchallengedgroup. Birds fed the combination of formulated yeast and B.licheniformis had similar (P>0.10) FCR and body weight gain to theunchallenged control group during d 0-21 and d 14-21 periods. Birds fedBMD had similar body weight gain to the unchallenged control during thed 0-21 period.

Necrotic enteritis lesion score was lower (P<0.05) in birds fed theformulated yeast and B. licheniformis combination treatment and BMDcompared to the challenged control birds. While birds fed BMD hadsimilar (P<0.10) necrotic enteritis lesion score as the unchallengedcontrol group. The unchallenged control, BMD and the combinationformulated yeast and B. licheniformis treatments had decreased (P<0.05)necrotic enteritis mortality compared to the challenged control group.The data suggests that both the combination formulated yeast and B.licheniformis product and BMD were effective in controlling C.perfringens disease challenge.

TABLE 22 Experimental treatments Conc. of E. maxima C. perfringens # ofTreatment Testing Product Testing Product Challenge Challenge Cages A NoAdditive None DOT¹ 14 None 8 B No Additive None DOT 14 DOT 19, 20, and21 8 C BMD 50.0 g/ton DOT 14 DOT 19, 20, and 21 8 D Formulated 0.5lb/ton + 3 × DOT 14 DOT 19, 20, and 21 8 yeast + B. licheniformis 10⁵cfu/g in final feed ¹Day of hatch

TABLE 23 Composition of the basal diet¹ Ingredients Kg/ton Corn, YellowGrain 556.29 Soybean Meal Dehulled, 377.55 Solvent Fat, Vegetable 25.55Dicalcium Phosphate 20.39 Calcium Carbonate 10.31 Salt, (NaCl) 4.61Methionine, MHA 3.08 L-LYSINE, 98.5% 0.82 Trace Mineral Premix 0.75Vitamin Premix 0.65 Total 1000.00 Calculated Nutrients Composition CrudeProtein, % 22.45 Fat, % 4.85 Crude Fiber, % 2.19 ME, kcal/Kg 3054.00Lysine, % 1.34 Methionine, % 0.63 Threonine, % 0.92 Calcium, % 0.92Phosphorus, % 0.45 Phosphorus, Available, % 0.67 Ca/P Ratio 2.04 Sodium,% 0.21 ¹Basal diet was prepared by Southern Poultry Research

TABLE 24 Summary 2018 SPRG NQ NE0716 Trial Treatment Non ChallengedChallenged Formulated Variable Control Control BMD yeast + B.licheniformis No. of Cages 8   8   8   8   No. of Birds 64    64   64    64    C. Perfringens − + + + Challenge Live performance, 0-21 dFeed 0.71 0.70 0.75 0.74 consumption, Kg Body weight  0.36^(a)  0.28^(b) 0.34^(a)  0.36^(a) gain, Kg Feed  1.97^(c)  2.49^(a)  2.21^(b) 2.06^(bc) Conversion Live performance, 14-21 d Feed 0.34 0.35 0.34 0.33consumption, Kg Body weight  0.18^(a)  0.13^(c)  0.15^(bc)  0.16^(ab)gain, Kg Feed  1.90^(c)  2.68^(a)  2.30^(b)  2.08^(bc) Conversion Liveperformance, 14-28 d Feed 0.84 1.01 0.78 0.83 consumption, Kg Bodyweight  0.44^(a)  0.30^(b)  0.38^(ab)  0.40^(a) gain, Kg Feed  1.90^(b) 3.37^(a)  2.05^(b)  2.08^(b) Conversion Live performance, 0-28 d Feed1.21 1.31 1.19 1.24 consumption, Kg Body weight 0.62 0.46 0.57 0.60gain, Kg Feed  1.95^(b)  2.84^(a)  2.08^(b)  2.06^(b) ConversionNecrotic enteritis 0.0^(c ) 1.1^(a ) 0.3^(c ) 0.6^(b ) lesion scoreNecrotic enteritis 0^(b )  17^(a )  3^(b )  2^(b )  related mortality, %^(a,b,c)Means within a row with no common superscripts differsignificantly (P < 0.05)

Example 9: Determine the Effect of Feeding Capsicum Product,Antimicrobial Clay and Bacillus licheniformis to Broilers Challengedwith C. perfringens

The study was conducted to evaluate the effects of Capsicum Product andAntimicrobial Clay with or without B. licheniformis in broilers raisedin pens on new litter (Table 25). A total of 1500 day-old male Ross×Rosschicks were obtained from a commercial hatchery. Chicks were vaccinatedfor Marek's and reovirus at the hatchery.

Experimental Diets

Feed consisted of non-medicated commercial-type broiler starter, grower,and finisher diets compounded according to meet or exceed NRC nutrientguidelines and contained feedstuffs commonly used in the United States(Table 26). Rations were fed ad libitum from date of chick arrival asfollows: Starter—DOT 0 until DOT 14, grower DOT 14 to DOT 28 andfinisher from DOT 28 to DOT 41 (study termination). Diets were fed ascrumbles (starter feed) or pellets (grower and finisher feed). Pelletingof diets (grower and finisher diets) were manufactured with a CaliforniaPellet mill (at 80° C.). Experimental treatment feeds were prepared froma basal feed formulation.

Animal Information

Upon arrival all birds were vaccinated (by spray cabinet) with acommercially approved coccidia vaccine at the normal recommended dosage.Only healthy chicks were used in this study.

Housing

Chicks were raised in 5×5 feet (1.5 m×1.5 m) floor pens (stockingdensity of 1.0 feet² per bird) in a solid-sided barn, with concretefloors, and under ambient temperature control. At placement, birds wereplaced on reused litter homogenized from a previous necrotic enteritisstudy. Each pen contained 1 (one) tube feeder and six (6) nippledrinkers.

Challenge Administration and Sample Collection

-   -   1. Necrotic Enteritis Challenge: The challenge model consisted        of coccidia from the DOT 0 vaccine and Clostridium perfringens        (using Clostridium perfringens strain #6 [Hofacre, et al.,        1998]) combination previously published by Hofacre, et al.        (1998).        -   A. Clostridium perfringens onto feed pans: Treatment feed            and water were withdrawn for two (2) to three (3) hours            prior to administration of Clostridium perfringens #6. A            measured amount of water (˜125 ml Clostridium perfringens #6            to 75 ml water) that was consumed within 30 minutes was used            for each pen. The Clostridium perfringens #6 fifteen (15)            hour culture at ˜1×10⁸ was added to this water and            thoroughly mixed and poured into the feed pan of birds in            each challenge pen. Clostridium perfringens #6 was dosed at            ˜1×10⁸ CFU/ml on DOT 20, 21, and 22 to birds in all            Treatment Groups.        -   B. Coccidia Challenge. Was from cycling of DOT 0 vaccination            and spreading E. maxima on feeders on DOT 15.    -   2. Necrotic Enteritis Scoring: DOT 23, three (3) birds per pen        were humanely euthanized, weighed necropsied and lesion scored        by Dr. Hofacre (Hofacre, 1998).        Lesion score 0=Normal        Lesion score 1=Slight mucus covering small intestine Lesion        score 2=Necrotic small intestine mucosa        Lesion score 3=Sloughed and blood small intestine mucosa and        contents

Body and Feed Weight

All birds were weighed by pen on DOT 0, 14, 28, and 41. The trial wasterminated on DOT 41. On DOT 14 remaining starter feed was removed,weighed, and replaced with grower feed to DOT 28. On DOT 28 remaininggrower feed was removed, weighed back, and replaced with finisher feedto DOT 41 (study termination).

Management

-   -   1. All birds were monitored for general flock condition,        temperature, lighting, water, feed, litter condition, and        unanticipated house conditions/events. Pens were checked daily        for mortality. A gross necropsy was performed on all dead or        culled birds to determine the bird sex and probable cause of        death.

Data was analyzed using one-way ANOVA by the MIXED procedure of SAS forthis complete randomized design. Pen served as the experimental unit.The statistical model included pen weight gain, feed consumption, feedconversion, percent NE related mortality and total percent mortality.All results were reported as least squares means. The significance levelchosen was α=0.05. Treatment effect was considered significant ifP<0.05, whereas values between 0.05≤P≤0.10 were considered asstatistical trends.

Summary

At 28 days of age birds fed the Antimicrobial Clay+B. licheniformistreatment combination and BMD had an increased (P<0.05) body weight gainand decreased (P<0.05) FCR compared to birds fed with no feed additive(Table 27). Broilers fed the combinations of Capsicum Product+B.licheniformis and Antimicrobial Clay+B. licheniformis and BMD had adecreased (P<0.05) FCR compared to the no feed additive control group at41 days of age. Body weight gain was increased (P<0.05) in birds fedAntimicrobial Clay+B. licheniformis and BMD compared to the control fedbirds at 41 days of age. Necrotic enteritis lesion score and necroticenteritis related mortality (Table 28) was numerically reduced in birdsfed the Capsicum Product+B. licheniformis, Antimicrobial Clay+B.licheniformis and BMD treatments compared to the no feed additivecontrol fed birds. These results suggest that feeding the combination ofCapsicum Product+B. licheniformis, Antimicrobial Clay+B. licheniformisand BMD were effective in controlling C. perfringens disease challenge.

TABLE 25 Experimental treatments Necrotic Enteritis Test Starter GrowerFinisher # of Treatment Challenge Product DOT¹ 0-13 DOT 14-35 DOT 35-41Pens A Yes No Additive None None None 10 B Yes BMD 50 g/ton 50 g/ton 50g/ton 10 C Yes Capsicum Product 0.3 lb/ton 0.3 lb/ton 0.3 lb/ton 10 DYes Capsicum Product + 0.3 lb/ton + 0.3 lb/ton + 0.3 lb/ton + 10 B.licheniformis 3 × 10⁵ 3 × 10⁵ 3 × 10⁵ cfu/g in cfu/g in cfu/g in finalfeed final feed final feed E Yes Antimicrobial Clay 4.0 lb/ton 4.0lb/ton 4.0 lb/ton 10 F Yes Antimicrobial Clay + 4.0 lb/ton + 4.0lb/ton + 4.0 lb/ton + 10 B. licheniformis 3 × 10⁵ 3 × 10⁵ 3 × 10⁵ cfu/gin cfu/g in cfu/g in final feed final feed final feed ¹Day of hatch

TABLE 26 Composition of the basal diets Ingredients, Kg/ton StarterGrower Finisher Corn, Yellow Grain 556.29 615.34 640.07 Soybean MealDehulled, 377.55 318.36 293.02 Solvent Fat, Vegetable 25.55 29.11 31.1Dicalcium Phosphate 20.39 18.94 17.86 Calcium Carbonate 10.31 9.85 9.18Salt, (NaCl) 4.61 4.39 4.41 Methionine, MHA 3.08 2.06 2.22 L-LYSINE,98.5% 0.82 0.70 0.89 Trace Mineral Premix 0.75 0.70 0.75 Vitamin Premix0.65 0.50 0.50 Total 1000.00 1000.00 1000.00 Calculated NutrientsComposition Crude Protein, % 22.45 20.0 19.0 Fat, % 4.85 5.35 5.61 CrudeFiber, % 2.19 2.12 2.10 ME, kcal/Kg 3054.00 3125.00 3165.00 Lysine, %1.34 1.16 1.11 Methionine, % 0.63 0.51 0.51 Threonine, % 0.92 0.82 0.77Calcium, % 0.92 0.85 0.80 Phosphorus, % 0.45 0.71 0.68 Phosphorus,Available, % 0.67 0.42 0.40 Ca/P Ratio, ratio 2.04 2.02 2.00 Sodium, %0.21 0.20 0.20

TABLE 27 Summary 2018 SPRG NQC072018-40 Trial (Example 5) TreatmentCapsicum Antimicrobial No Feed Capsicum Product + B. AntimicrobialClay + B. Variable Additive BMD Product licheniformis Clay licheniformisNo. of Pens 10 10 10 10 10 10 No. of Birds 250 250 250 250 250 250 Liveperformance, 14 d Feed consumption, Kg 0.44 0.44 0.45 0.43 0.48 0.47Body weight gain, Kg 0.33^(bc) 0.33^(abc) 0.35^(a) 0.32^(c) 0.34^(abc)0.35^(ab) Feed Conversion 1.35^(ab) 1.35^(ab) 1.30^(b) 1.35^(ab)1.40^(a) 1.35^(ab) Live performance, 28 d Feed consumption, Kg 1.87 1.991.90 1.88 1.91 1.94 Body weight gain, Kg 1.15^(c) 1.26^(a) 1.17^(c)1.18^(bc) 1.15^(c) 1.23^(ab) Feed Conversion 1.63^(ab) 1.58^(c)1.62^(abc) 1.59^(bc) 1.66^(a) 1.58^(c) Live performance, 41 d Feedconsumption, Kg 3.95 4.08 3.88 3.89 3.92 3.99 Body weight gain, Kg2.34^(c) 2.47^(a) 2.34^(c) 2.37^(bc) 2.36^(c) 2.45^(ab) Feed Conversion1.69^(a) 1.65^(b) 1.66^(ab) 1.64^(b) 1.66^(ab) 1.63^(b) Total mortality,% 6.0^(ab) 4.4^(b) 12.0^(a) 8.4^(ab) 11.6^(a) 8.0^(ab) ^(a,b,c)Meanswithin a row with no common superscripts differ significantly (P < 0.05)

TABLE 28 2018 SPRG NQC072018-40 Trial (Example 5) Clinical necroticenteritis Percent Average NE Treatment NE Mortality Lesion Score No FeedAdditive 2.40^(b) 0.53 BMD 1.20^(b) 0.43 Capsicum Product 7.60^(a) 0.40Capsicum Product + B. licheniformis 1.60^(b) 0.33 Antimicrobial Clay4.80^(ab) 0.53 Antimicrobial Clay + B. licheniformis 1.60^(b) 0.50

What is claimed is:
 1. A feed composition for non-human animalscomprising a basal animal diet supplemented with a combination of activeingredients selected from capsicum product and a DFM; antimicrobial clayand a DFM; formulated yeast and a DFM; capsicum product, formulatedyeast, and a DFM, and capsicum product and formulated yeast.
 2. Thecomposition of claim 1, wherein the DFM is Bacillus licheniformis. 3.The composition of claim 2, wherein the Bacillus licheniformis in thefeed composition is at a concentration of about 1×10⁴ cfu/g to about1×10⁶ cfu/g feed.
 4. The composition of claim 1, wherein the capsicumproduct in the feed composition is at a concentration of about 0.1lb/ton to about 0.5 lb/ton feed.
 5. The composition of claim 1, whereinthe capsicum product is selected from the group consisting of acapsaicinoid, a vanilloid, capsicum, macerated hot peppers, ground hotpeppers, hot pepper extract, capsaicin-containing plant materials,encapsulated ground peppers, a coated capsaicin product, andcombinations of any thereof.
 6. The composition of claim 1, wherein thecapsicum product is encapsulated ground peppers.
 7. The composition ofclaim 6, wherein the capsicum product comprises about 45% to about 55%ground peppers.
 8. The composition of claim 6, wherein the capsicumproduct comprises about 0.4% to about 0.6% capsaicinoids.
 9. Thecomposition of claim 1, wherein the formulated yeast comprises about80-100% yeast extract.
 10. The composition of claim 9, wherein theformulated yeast in the feed composition is at a concentration of about0.3 lb/ton of feed to about 0.7 lb/ton of feed.
 11. The composition ofclaim 1, wherein the clay is mined clay.
 12. The composition of claim11, wherein the antimicrobial clay in the feed composition is at aconcentration of about 2 to about 6 lb/ton feed.
 13. The composition ofclaim 1, wherein the composition comprises formulated yeast, capsicumproduct, and DFM.
 14. The composition of claim 13, wherein the DFM isBacillus licheniformis.
 15. The composition of claim 14, wherein theformulated yeast in the feed composition is at a concentration of about0.3 lb/ton to about 0.7 lb/ton feed, the capsicum product in the feedcomposition is at a concentration of about 0.1 lb/ton to about 0.5lb/ton feed, and the Bacillus licheniformis in the feed composition isat a concentration of about 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed. 16.The composition of claim 1, wherein the composition comprises capsicumproduct and DFM.
 17. The composition of claim 16, wherein DFM isBacillus licheniformis.
 18. The composition of claim 17, wherein thecapsicum product in the feed composition is at a concentration of about0.1 lb/ton to about 0.5 lb/ton feed and the Bacillus licheniformis inthe feed composition is at a concentration of about 1×10⁴ cfu/g to about1×10⁶ cfu/g feed.
 19. The composition of claim 1, wherein thecomposition comprises antimicrobial clay and DFM.
 20. The composition ofclaim 19, wherein DFM is Bacillus licheniformis.
 21. The composition ofclaim 20, wherein the antimicrobial clay in the feed composition is at aconcentration of about 2 lb/ton to about 6 lb/ton feed and the Bacilluslicheniformis in the feed composition is at a concentration of about1×10⁴ cfu/g to about 1×10⁶ cfu/g feed.
 22. The composition of claim 1,wherein the composition comprises formulated yeast, and DFM.
 23. Thecomposition of claim 22, wherein DFM is Bacillus licheniformis.
 24. Thecomposition of claim 23, wherein the formulated yeast in the feedcomposition is at a concentration of about 0.3 lb/ton to about 0.7lb/ton feed, and the Bacillus licheniformis in the feed composition isat a concentration of about 1×10⁴ cfu/g to about 1×10⁶ cfu/g feed. 25.The composition of claim 1, wherein the composition comprises formulatedyeast, and capsicum product.
 26. The composition of claim 25, whereinthe formulated yeast in the feed composition is at a concentration ofabout 0.3 lb/ton to about 0.7 lb/ton feed, and the capsicum product inthe feed composition is at a concentration of about 0.1 lb/ton to about0.5 lb/ton feed.
 27. A feed composition for non-human animals comprisinga basal animal diet supplemented with capsicum product and Bacilluslicheniformis.
 28. A feed composition for non-human animals comprising abasal animal diet supplemented with antimicrobial clay and Bacilluslicheniformis.
 29. A feed composition for non-human animals comprising abasal animal diet supplemented with formulated yeast and Bacilluslicheniformis.
 30. A feed composition for non-human animals comprising abasal animal diet supplemented with capsicum product, formulated yeast,and Bacillus licheniformis.
 31. A feed composition for non-human animalscomprising a basal animal diet supplemented with capsicum product andformulated yeast.
 32. A method for improving health in non-humananimals, the method comprising feeding the non-human animal a feedcomposition of claim
 1. 33. The method of claim 32, wherein improvingthe health of a non-human animal is selected from reducing incidence ofdiarrhea, reducing number of days of diarrhea, decreasing mortality,improving intestinal health, reducing microbial pathogens in theintestinal tract of the animal, decreasing cytokine panel measuringTNF-alpha, and combinations thereof.
 34. The method of claim 33, whereinthe non-human animal is a chicken.
 35. The method of claim 34, whereinimproving the health of the chicken comprises reducing the impact of C.perfringens in broilers.
 36. A method for improving performance ofnon-human animals, the method comprising feeding the non-human animal afeed composition of claim
 1. 37. The method of claim 36, whereinimproving performance is selected from increase in body weight gain,feed conversion rate (FCR), feed intake, average daily weight gain(ADG), an increase in average daily food intake (ADFI), an improvedoverall body weight, and ratio of F/G.
 38. The method of claim 36,wherein the non-human animal is a chicken.
 39. A method for improvingquality of a product derived from a non-human animal, the methodcomprising feeding the non-human animal a feed composition of claim 1.40. The method of claim 39, wherein improving the quality of a productderived from a non-human animal is selected from improving breast meatyield, reducing product variability, reducing impact on consumer appealdue to poor color, poor texture, and drip loss in retail display,reducing organoleptic qualities in cooked chicken, including incidenceof woody breast.
 41. The method of claim 39, wherein the non-humananimal is a chicken.